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An English Medium Co. Ed. Senior

Secondary School

Investigatory

Project

On

SUBMITTE B!"SUBMITTE B!"

SUBMITTE TO"SUBMITTE TO"

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Su#hag Singh Mr.Su#hag Singh Mr.

Sandee$ %ulshesthraSandee$ %ulshesthra

&II Sci. B&II Sci. B

'(.O. Biology)'(.O. Biology)

A*no+ledgementI am over+helmed in all hum#leness and grate,ulness to ac*no+ledge

my de$th to all those +ho have hel$ed me to $ut these ideas- +ell

a#ove the level o, sim$licity and into something concrete.I +ould li*e to e$ress my s$ecial than*s o, gratitude to my #iology

teacher- Mr. Sandeep Kulshesthra as +ell as our Princi$al Mrs. Nidhi

Bhatia  +ho gave me the golden o$$ortunity to do this +onder,ul

$roject on the to$ic /A$$lications o, Biotechnology0- +hich also hel$ed

me in doing a lot o, research and I came to *no+ a#out so many ne+

things. I am really than*,ul to them.

Any attem$t at any level can1t #e satis,actorily com$leted +ithout the

su$$ort and guidance o, my Parents and Friends +ho hel$ed me a lotin gathering di2erent in,ormation- collecting data and guiding me ,rom

time to time in ma*ing this $roject- des$ite o, their #usy schedules- they

gave me di2erent ideas in ma*ing this $roject uni3ue. I am than*,ul to

them too.

I am ma*ing this $roject not only ,or mar*s #ut to also increase my

*no+ledge...

 Than*ing you

Subhag SinghXII Sci. B

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Certi4cate This is to certi,y that SUBHAG SINGH o, class

XII SCI.B o, GAN !""P SHIKSHA BHA#A$I

has success,ully com$leted the investigatory

$roject on the to$ic %APP&ICA$I'NS 'FBI'$"CHN'&'G(  under the guidance o, 

M#. SAN!""P KU&SH"S$H#A  '(.O..

Biology) during the session 567897: in the

$artial ,ul4lment o, Biology Practical

Eamination conducted #y C"N$#A& B'A#!

'F S"C'N!A# "!UCA$I'N )AISSC"*.

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 ;;;;;;;;;;;;;;;;;;; ;;;;;;;;;;;;;;;;;;; Mr. Sandee$ %ulshesthra Eternal

Eaminer'(.O. Biology) 'C.B.S.E)

Introduction+hat is Bi,techn,l,g- 

Bi,techn,l,g- is the use o, living systems and organisms to develo$ orma*e $roducts- or <any technological a$$lication that uses #iologicalsystems- living organisms or derivatives thereo,- to ma*e or modi,y$roducts or $rocesses ,or s$eci4c use.

 At its sim$lest- #iotechnology

is technology #ased on #iology 9#iotechnology harnesses

cellular and #io molecular

$rocesses to develo$

technologies and

$roducts that hel$

im$rove our lives and the

health o, our $lanet.

=e have used the #iological$rocesses o, microorganisms ,or

more than :-666 years to ma*e

use,ul ,ood $roducts- such as #read and cheese- and to $reserve dairy

$roducts.

  Modern #iotechnology $rovides #rea*through $roducts and

technologies to com#at de#ilitating and rare diseases- reduce our

environmental ,oot$rint- ,eed the hungry- useless and cleaner energy-

and have sa,er- cleaner and more e>cient industrial manu,acturing$rocesses.

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Biotech is hel$ing to heal the +orld #y harnessing nature?s o+n tool#oand using our o+n genetic ma*eu$ to heal and guide lines o, research#y"

• @educing rates o, in,ectious disease

• Saving millions o, children?s lives

• Changing the odds o, serious- li,e9threatening conditions a2ectingmillions around the +orld

•  Tailoring treatments to individuals to minimie health ris*s andside e2ects

• Creating more $recise tools ,or disease detection

• Com#ating serious illnesses and everyday threats con,ronting thedevelo$ing +orld.

(istory

 Throughout the history o, agriculture- ,armers have inadvertently

altered the genetics o, their cro$s through introducing them to ne+

environments and #reeding them +ith other $lants 9 one o, the 4rst

,orms o, #iotechnology.

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 These $rocesses also +ere included in early /er0entati,n ,/ beer.

In #re+ing- malted grains 'containing enymes) convert starch ,rom

grains into sugar and then adding

s$eci4c yeasts to $roduce #eer. In

this $rocess- car#ohydrates in the

grains +ere #ro*en do+n intoalcohols such as ethanol. ater

other cultures $roduced the

$rocess o, lactic acid

,ermentation +hich allo+ed the

,ermentation and $reservation o, 

other ,orms o, ,ood- such as s,-

sauce. ermentation +as also

used in this time $eriod to $roduce lea1ened bread. Although the$rocess o, ,ermentation +as not ,ully understood until ouis Pasteur?s

+or* in 7D8- it is still the 4rst use o, #iotechnology to convert a ,ood

source into another ,orm.

or thousands o, years- humans have used selective #reeding to

im$rove $roduction o, cro$s and livestoc* to use them ,or ,ood. In

selective #reeding- organisms +ith desira#le characteristics are mated

to $roduce o2s$ring +ith the same characteristics. or eam$le- this

techni3ue +as used +ith corn to $roduce the largest and s+eetestcro$s.

Biotechnology has also led to the develo$ment o, anti#iotics. In

7F5D- Aleander leming discovered the mould Penicilliu0. (is +or*

led to the $uri4cation o, the anti#iotic com$ound ,ormed #y the mould

#y (o+ard lorey- Ernst Boris Chain and Gorman (eatley 9 to ,orm +hat

+e today *no+ as $enicillin. In 7FH6- $enicillin #ecame availa#le ,or

medicinal use to treat #acterial in,ections in humans.

 The 4eld o, modern #iotechnology is generally thought o, as having

#een #orn in 7F7 +hen Paul Berg?s e$eriments in gene splicing had

early success. (er#ert =. Boyer and Stanley G. Cohen signi4cantly

advanced the ne+ technology in 7F5 #y trans,erring genetic material

into a #acterium- such that the im$orted material +ould #e re$roduced.

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Biotechnology in

AgricultureGeneticall- M,di2ed Cr,ps

Geneticall- 0,di2ed cr,ps or /M

cro$s0 or /#iotech cro$s0 are $lants used

in agriculture- the GA o, +hich has #een

modi4ed +ith genetic

engineering techni3ues. In most cases

the aim is to introduce a ne+ trait to the

$lant +hich does not occur naturally in

the s$ecies. Eam$les in ,ood cro$s

include resistance to certain $ests- diseases- stress,ul environmental

conditions- resistance to chemical treatments- reduction o, s$oilage- or

im$roving the nutrient $ro4le o, the cro$. Eam$les in non9,ood cro$s

include $roduction o, $harmaceutical agents- #io ,uels- and other

industrially use,ul goods- 

as +ell as ,or #ioremediation.

Plants and cro$s +ith M traits have #een tested more than any other

cro$sJ+ith no credi#le evidence o, harm to humans or animals. In ,act-

seeds +ith M traits have #een tested more than any other cro$s in the

history o, agriculture K +ith no credi#le evidence o, harm to humans or

animals.

overnmental regulatory agencies- scienti4c organiations and leading

health associations +orld+ide agree that ,ood gro+n ,rom GM cr,ps issa/e t, eat. The =orld (ealth Organiation- the American Medical

Association- the U.S. Gational Academy o, Sciences- the British @oyal

Society- among others that have eamined the evidence- all come to the

same conclusion" consuming ,oods containing ingredients derived ,rom

M cro$s is sa,e to eat and no ris*ier than consuming the same ,oods

containing ingredients ,rom cro$ $lants modi4ed #y conventional $lant

im$rovement techni3ues. enetic modi4cations have"

7. Made cro$s more tolerant to a#iotic stresses 'cold- drought- salt-

heat).

5. @educed reliance on chemical $esticides '$est resistant cro$s).

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L. (el$ed to reduce $ost harvest losses enhanced the nutritional

value o, the ,ood.

#NA Inter/erence )#NAi*

@GA inter,erence '@GAi) is a method o, #loc*ing gene ,unction #y

inserting short se3uences o, ri#onucleic acid '@GA) that match $art o, 

the target gene1s se3uence- thus no $roteins are $roduced. @GAi has the

$otential to #ecome a $o+er,ul thera$eutic a$$roach to+ard targeted

and $ersonalied medicine. @GAi has $rovided a +ay to control $estsand diseases- introduce novel $lant traits and increase cro$ yield. Using

@GAi- scientists have develo$ed novel cro$s such as nicotine9,ree

to#acco- non9allergenic $eanuts- deca2einated co2ee- and nutrient

,orti4ed maie among many others.

Mechanism o, @GA inter,erences as understood is that it comes into $lay

+hen a dou#le stranded @GA is introduced either naturally or arti4cially

in a cell. An endo ri#onuclease enyme cleaves the long ds@GA into

small $ieces o, @GA. The small $ieces could #e mi @GA or si @GA

de$ending u$on the origin o, long ds@GA i.e. endogenous or eogenous

res$ectively. A dou#le

stranded @GA may #e

generated #y either @GA

de$endent @GA $olymerase

or #idirectional transcri$tion

o, trans$osa#le elements or

$hysically introduced.

 There are several

o$$ortunities ,or the

a$$lications o, @GAi in cro$

science ,or its im$rovement

such as stress tolerance and

enhanced nutritional level.This *noc*do+n technology may #e use,ul in

inducing early No+ering- delayed ri$ening- delayed senescence-

#rea*ing dormancy- stress9,ree $lants- overcoming sel,9sterility- etc. 

@GA inter,erence '@GAi) has recently #een demonstrated in $lant

$arasitic nematodes. It is a $otentially $o+er,ul investigative tool ,or the

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genome9+ide identi4cation o, gene ,unction that should hel$ im$rove

our understanding o, $lant $arasitic nematodes. @GAi should hel$

identi,y gene and- hence- $rotein targets ,or nematode control

strategies. Pros$ects ,or novel resistance de$end on the $lant

generating an e2ective ,orm o, dou#le9stranded @GA in the a#sence o, 

an endogenous target gene +ithout detriment to itsel,. These @GAmolecules must then #ecome availa#le to the nematode and #e ca$a#le

o, ingestion via its ,eeding tu#e. I, these re3uirements can #e met- cro$

resistance could #e achieved #y a $lant delivering a ds@GA that targets

a nematode gene and induces a lethal or highly damaging @GAi e2ect

on the $arasite.

Bt t,3in

A $rotein that is toic to che+ing insects and is $roduced #y the soil#acterium Bacillus thuringiensis and has long #een used as a #iologicalpesticide. By means o, genetic engineering- the genes ,or Bt toin can#e isolated ,rom Bacillus thuringiensis and trans,erred to $lants.

Bacillus thuringiensis )Bt* is a #acteria that $roduces $roteins +hichare toic to insects. But etreme toicity comes at no sur$rise. It1s in thesame ,amily o, #acteria as B. anthracis- +hich causes anthra- and B.cereus- +hich causes ,ood $oisoning.

 The Bt  toin dissolve in the high $( insect gut and #ecome active. The

toins then attac* the gut cells o, the insect- $unching holes in thelining. The Bt  s$ores s$ills out o, the gut and germinate in the insectcausing death +ithin a cou$le days.

Even though the toin does not *ill the insect immediately- treated $lant$arts +ill not #e damaged #ecause the insect sto$s ,eeding +ithinhours. Bt  s$ores do not s$read to other insects or cause diseaseout#rea*s on their o+n.

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7. Insect eats Bt  crystals and s$ores.

 5. The toin #inds to s$eci4crece$tors in the gut and the insectssto$s eating.

L. The crystals cause the gut +all to#rea* do+n- allo+ing s$ores andnormal gut #acteria to enter the #ody.

H. The insect dies as s$ores and gut#acteria $roli,erate in the #ody.

Bt  action is very s$eci4c. i2erent strains o, Bt  are s$eci4c to di2erentrece$tors in insect gut +all. Bt  toicity de$ends on recogniingrece$tors- damage to the gut #y the toin occurs u$on #inding to arece$tor. Each insect s$ecies $ossesses di2erent ty$es o, rece$tors that+ill match only certain toin $roteins- li*e a loc* to a *ey.

It is #ecause o, this that ,armers have to #e care,ul to match the target$est s$ecies +ith a $articular Bt  toin $rotein +hich is s$eci4c ,or thatinsect. This also hel$s the #eni4cal insects #ecause they +ill usually not#e harmed #y that $articular strain o, Bt .

Bt C,tt,n

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Bt c,tt,n is a genetically modi4ed organism 'MO) cotton variety-

+hich $roduces an insecticide to #oll+orm. Strains o, the

#acterium Bacillus thuringiensis $roduce over 566 di2erent Bt toins-

each harm,ul to di2erent insects. Most nota#ly-

Bt toins are insecticidal to the larvae o, moths

and #utterNies- #eetles- cotton #oll+orms andghtu Nies #ut are harmless to other ,orms o, li,e.

 The gene coding ,or Bt toin has #een inserted

into cotton as a transgene- causing it to $roduce

this natural insecticide in its tissues. In many

regions- the main $ests in commercial cotton

are le$ido$teran larvae- +hich are *illed #y the

Bt $rotein in thegenetically modi4ed cotton they

eat. This eliminates the need to use largeamounts o, #road9s$ectrum insecticides to *ill le$ido$teran $ests. This

s$ares natural insect $redators in the ,arm ecology and ,urther

contri#utes to non insecticide $est management. 

Bt cotton is ine2ective against many cotton $ests such as $lant

#ugs- stin* #ugs- and a$hids de$ending on circumstances it may #e

desira#le to use insecticides in $revention. A 566: study done

#y Cornell researchers- the Center ,or

Chinese Agricultural Policy and the ChineseAcademy o, Science on Bt cotton ,arming in

China ,ound that a,ter seven years these

secondary $ests that +ere normally

controlled #y $esticide had increased-

necessitating the use o, $esticides at similar

levels to non9Bt cotton and causing less

$ro4t ,or ,armers #ecause o, the etra

e$ense o, M seeds.

Mechanism:

Bt cotton +as created through the addition o, genes encoding toin

crystals in the Cry grou$ o, endotoin. =hen insects attac* and eat the

cotton $lant the Cry toins are dissolved due to the high $( level o, the

insects stomach. The dissolved and activated Cry molecules #ond to

cadherin9li*e $roteins on cells com$rising the #rush #order

molecules. The e$ithelium o, the #rush #order mem#ranes se$arates

the #ody cavity ,rom the gut +hilst allo+ing access ,or nutrients. The

Cry toin molecules attach themselves to s$eci4c locations on the

cadherin9li*e $roteins $resent on the e$ithelial cells o, the midge and

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ion channels are ,ormed +hich allo+ the No+ o, $otassium. @egulation

o, $otassium concentration is essential and- i, le,t unchec*ed- causes

death o, cells. ue to the ,ormation o, Cry ion channels su>cient

regulation o, $otassium ions is lost and results in the death o, e$ithelial

cells. The death o, such cells creates ga$s in the #rush #order

mem#rane. 

 Advantages:

Bt cotton has several advantages over non Bt cotton. The im$ortant

advantages o, Bt cotton are #rieNy "

• Increases yield o, cotton due to e2ective control o, three ty$es o, #oll+orms- vi. American- S$otted and Pin* #oll+orms.

• Insects #elonged to e$ido$tera 'Boll+orms) are sensitive tocrystalline endotoic $rotein $roduced #y Bt gene +hich in turn$rotects cotton ,rom #oll+orms.

• @eduction in $esticide use in the cultivation o, Bt cotton in +hich#oll+orms are major $ests.

• @eduction in the cost o, cultivation and lo+er ,arming ris*s.

• @eduction in environmental $ollution #y the use o, insecticidesrarely. 

• Bt cotton ehi#it genetic resistance or in#uilt resistance +hich is a$ermanent ty$e o, resistance and not a2ected #y environmental

,actors. Thus $rotects cro$ ,rom #oll+orms.

• Bt cotton is eco,riendly and does not have adverse e2ect on$arasites- $redators- #ene4cial insecticides and organisms $resent insoil.

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• It $romotesmulti$lication o,  $arasites and $redators+hich hel$ incontrolling the

#oll+orms #y ,eedingon larvae and eggs o, #oll+orm.

• Go health haardsdue to rare use o, insecticides.

• Bt cotton are early in maturing as com$ared to non Bt cotton.

Disadvantages:Bt cotton has some limitations

• (igh cost o, Bt cotton seeds as com$ared to non Bt cotton seeds.

• E2ectiveness u$ to 756 days- a,ter that the toin $roducinge>ciency o, the Bt gene drastically reduces.

• Ine2ective against suc*ing $ests li*e jassids- a$hids- +hiteNy etc.

Bt cotton in India:

Bt cotton is su$$lied in India?s Maharashtra state #y the agri9

#iotechnology com$any- Mahyco- as the distri#utor.

 The use o, Bt cotton in India has gro+n e$onentially since its

introduction. @ecently India has #ecome the num#er one glo#al e$orter

o, cotton and the second largest cotton $roducer in the +orld. India has

#red Bt9cotton varieties such as Bikaneri Nerma and hy#rids such as

G((9HH- setting u$ India to #ene4t no+ and +ell into the ,uture.

India1s success has #een su#ject to scrutiny. Monsanto?s seeds are

e$ensive and lose vigour a,ter one generation- $rom$ting the Indian

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Council o, Agricultural @esearch to develo$ a chea$er Bt cotton variety

+ith seeds that could #e reused. The cotton incor$orated the cry7Ac

gene ,rom the soil #acterium Bacillus thuringiensis 'Bt)- ma*ing the

cotton toic to #oll+orms. In $arts o, India cases o, ac3uired resistance

against Bt cotton have occurred.

 The state o, Maharashtra #anned the sale and distri#ution o, Bt cotton

in 5675- to $romote local Indian seeds- +hich demand less +ater-

,ertiliers and $esticide in$ut- #ut li,ted the #an in 567L.

India a$$roved Bt cotton in 5665 no+ it accounts ,or F5 o, all Indian

cotton. Average nation+ide cotton yields +ent ,rom L65 *gQha in the

5665QL season to a $rojected HD7 *gQha in 5677Q75 J u$ 8F.L overall.

 This chart sho+s the trends in yields- +hich too* o2 a,ter Bt +as

introduced in 5665. The gra$hs also sho+ that J and here comes ugly

,actJ in the last H years- as Bt has risen ,rom : to F5 o, India1s

cotton- yields have dro$$ed steadily.

Biotechnology inMedicine

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Geneticall- "ngineered Insulin )Hu0ulin*

Insulin  is a $e$tide hormone $roduced

#y #eta cells in the $ancreas o, various

organisms including human #eings. It

regulates

the 0etab,lis0 ,/ carb,h-drates an

d ,ats #y $romoting the a#sor$tion

o, glucose ,rom the #lood to s*eletal

muscles and ,at tissue and #y causing

,at to #e stored rather than used ,or energy. Insulin also inhi#its the

$roduction o, glucose #y the liver.

Ece$t in the $resence o, the meta#olic disorder dia#etes

mellitus and meta#olic syndrome- insulin is $rovided +ithin the #ody in

a constant $ro$ortion to remove ecess glucose ,rom the #lood- +hich

other+ise +ould #e toic. =hen #lood glucose levels ,all #elo+ a certain

level- the #ody #egins to use stored glucose as an energy source

through glycogenolysis- +hich #rea*s do+n the glycogen stored in the

liver and muscles into glucose- +hich can then #e utilied as an energy

source. As a central meta#olic control mechanism- its status is also used

as a control signal to other #ody systems 'such as amino acid u$ta*e #y#ody cells). In addition- it has several other ana#olic e2ects throughout

the #ody. +hen c,ntr,l ,/ insulin le1els /ails4 diabetes

0ellitus can result.

Structure:

Insulin is com$osed o, t+o

di2erent ty$es o, $e$tidechains. Chain A has 57 amino

acids and Chain B has L6 amino

acids. Both chains contain al$ha

helices #ut no #eta strands.

 There are L conserved disul2de

bridges +hich hel$ *ee$ the

t+o chains together. Insulin can

also ,orm dimers in solution due

to the hydrogen #onding #et+een the B chains. The dimers can ,urther

interact to ,orm heamers due to interaction #et+een hydro$ho#ic

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sur,aces. This scene highlights the hydro$ho#ic and $olar $arts o, an

insulin monomer at a $( o, .

A num#er o, insulin variants have #een made to ,avor either themonomeric or heameric ,orm. eletion o, the 4ve C terminal residues

o, the B chain creates a monomer only ,orm. This $ortion o, the B chain

is involved in hydrogen #onds #et+een the B chain o, one monomer and

the A and B chain o, another monomer . 

Need of Genetically Engineered Insulin:

 The original ,orm o, the +onder cure ,or dia#etes- these +ere once the

only ty$e o, insulin availa#le- #ut are no+ rarely used. Ani0al insulin

+as originally made

,rom ground9u$

animal $ancreas

tissue- and then later

+as etracted ,rom

healthy animals

'slaughtered $igs co+s). The

meta#olism o, co+s and $igs +as close enough to human meta#olism

that their animal insulin also +or*ed +ell in human #odies. Bee, insulin

has L di2erences ,rom human $or* insulin has 7 di2erence ,rom

human. The use o, a miture o, #ee, and $or* insulin +as also $ossi#le.

 It has #een sho+n that human insulin is less immunogenic than animal

insulin. Porcine insulin is most similar to human insulin. The $rimary

amino acid se3uences o, #ovine and $orcine insulin di2er ,rom that o, human insulin #y three and one amino acid- res$ectively. This greater

dissimilarity #et+een human and #ovine insulin has #een $ostulated to

#e the e$lanation ,or the greater antigenicity o, #ovine insulin as

com$ared +ith $orcine insulin

One o, the pr,ble0s 5ith ani0al insulin 5as antib,d- issues. The

#ody identi4es them and tries to reject them. Por* insulin di2ers #y 7

amino acid and #ee, insulin #y L amino acids- so the #ody?s immune

system can sometimes recognie them as ,oreign. Immunologicalcom$lications o, insulin thera$y have #een evident since animal insulin

#ecame availa#le ,or the treatment o, dia#etes mellitus in 7F55. In

insulin9allergic $atients treated +ith conventional insulin $re$arations-

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the insulin9s$eci4c IgE values are o,ten 769 to 569,old higher than in

$atients +ithout allergy. It has #een sho+n that human insulin is less

immunogenic than animal insulin. Porcine

insulin is most similar to human insulin. Cross9

reactivity #et+een human insulin and insulin

o, animal origin has #een re$orted. A major$ro#lem is the cross9reactivity that occurs

#et+een anti9insulin anti#odies and the

various animal and human insulin $re$arations

in $atients $resenting +ith allergy to animal

insulin.

 The usage o, ani0al insulin has s, greatl- declined in 0,dern

ti0es that they have largely #een +ithdra+n ,rom the mar*et. Ge+ly

diagnosed dia#etics are ty$ically given synthesied or Geneticall-

"ngineered hu0an insulin.

 

What is !roinsulin"#

Pr,insulin is the $rohormone $recursor to insulin made in the #etacells o, the islets o, angerhans- s$ecialied regions o, the $ancreas.

Proinsulin is synthesied on

mem#rane associated

ri#osomes ,ound on the rough

endo$lasmic reticulum- +here it

is ,olded and its disul4de

#onds are oidied. It is then

trans$orted to the olgi

a$$aratus +here it is $ac*aged

into secretory vesicles- and

+here it is $rocessed #y a series

o, $roteases to ,orm

mature insulin. Mature insulin has L8 ,e+er amino acids H are removed

altogether- and the remaining L7 ,orm the C9$e$tide. The C9$e$tide is

a#stracted ,rom the center o, the $roinsulin se3uence the t+o other

ends 'the B chain and A chain) remain connected #y disul4de #onds.

 =hen insulin +as originally $uri4ed ,rom #ovine or $orcine $ancreata-

all the $roinsulin +as not ,ully removed.RLRH =hen some $eo$le used

these insulins- the $roinsulin may have caused the #ody to react +ith a

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rash- to resist the insulin- or even to ma*e dents or lum$s in the s*in at

the $lace +here the insulin +as injected. This can #e descri#ed as

an iatrogenic injury due to slight di2erences #et+een the $roinsulin o, 

di2erent s$ecies. Since the late 7F6s- +hen highly

$uri4ed $orcine  insulin +as introduced- and the level o, insulin $urity

reached FF- this ceased to #e a signi4cant clinical issue. $he 0ainchallenge /,r pr,ducti,n ,/ insulin using r!NA techni6ues 5as

getting insulin asse0bled int, 0ature /,r0.

$umulin:

 Hu0ulin  +as the 4rst medication $roduced using modern genetic

engineering techni3ues in +hich actual human GA is inserted into ahost cell 'E. coli in this case). Biosynthetic <human< insulin is no+

manu,actured ,or +ides$read clinical use using genetic engineering

techni3ues using recom#inant GA technology- +hich the

manu,acturers claim reduces the $resence o, many im$urities- although

there is no clinical evidence to su#stantiate this claim. "li

&ill- 0ar7eted the 2rst arti2cial insulin4 Hu0ulin4 in 89:;.

(umulin $roduction method is as ,ollo+s"

7. GA coding ,or A and B $oly$e$tide chains o, insulin are

chemically synthesised a in the la#. Sity three nucleotides are

se3uenced to $roduce A chain o, insulin and ninety nucleotide long

GA designed to $roduce B chain o, insulin- $lus terminator codon

is added at the end o, each chain se3uence. Anti9codon ,or

methionine is added at the #eginning o, the se3uence to

distinguish humulin ,rom the other #acterial $roteins.

5. Chemically synthesied A and B chain GA se3uence are inserted

into one o, the mar*er gene +hich are $resent in the $lasmid

vector. enes are inserted into the $lasmid +ith the hel$ o, 

enymes *no+n as endonuclease and ligase.

L. The vector $lasmids +ith the insulin gene are then introduced into

the E. coli #acterial cell. These cells are then allo+ed to re$licate

#y mitosis- along +ith the #acterial cell recom#inant $lasmid also

gets re$licated $roducing the human insulin.

H. A and B $oly$e$tide chains o, insulin are then etracted and

$uri4ed ,rom the ,omenters in the la#. (igh9Per,ormance i3uid

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Chromatogra$hy '(PC) is used to get 766 $ure humulin ,rom

the miture o, $roteins.

8. The A and B $oly$e$tide chains o, insulin are mied together and

connected +ith each other #y disul$hide #ond- ,orming the

(umulin or synthetic human insulin.

 Advantages % Disadvantages of $umulin:

(umulin is the one and only human $rotein $roduced in the #acteria

+ith identical chemical structure to that o, the natural human insulin.

Administration o, humulin reduces the $ossi#ility o, anti#ody $roduction

and inNammatory res$onsein dia#etic $atients. Major

di>culty is the etraction o, 

humulin ,rom a miture o, 

host $roteins $resent in the

,ermentation #roth.

Go+ days to overcome this

etraction $ro#lem synthetic

human insulin are $roduced

in the yeast cell instead o, E. coli using the same $rocedure. As yeast is

Eu*aryotes they secrete the +hole humulin molecule +ith $er,ect three

dimensional structures- reducing the need ,or com$le and time

consuming $uri4cation methods.

Go+ most o, the dia#etic $atients are treated +ith synthetic human

insulin. Small grou$ o, $atients claim that e$isodes o, hy$erglycaemic

com$lications have #een increased a,ter shi,ting ,rom animal origin

insulin to humulin. Go study till date sho+s the di2erence #et+een the

,re3uency o, hy$erglycaemic com$lications in $atient using humulin

'synthetic human insulin) and animal origin insulin.

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Gene $herap-

Gene therap- is the thera$eutic delivery o, nucleic acid $olymers into

a $atient?s cells as a drug to treat disease. ene thera$y is an

e$erimental techni3ue that uses genes to treat or $revent disease. In

the ,uture- this techni3uemay allo+ doctors to treat a

disorder #y inserting a gene

into a $atient1s cells instead

o, using drugs or surgery.

@esearchers are testing

several a$$roaches to gene

thera$y- including"

• @e$lacing a mutated

gene that causes

disease +ith a healthy

co$y o, the gene.

• Inactivating- or /*noc*ing out-0 a mutated gene that is ,unctioning

im$ro$erly.

•

Introducing a ne+ gene into the #ody to hel$ 4ght a disease.

Although gene thera$y is a $romising treatment o$tion ,or a num#er o, 

diseases 'including inherited disorders- some ty$es o, cancer- and

certain viral in,ections)- the techni3ue remains ris*y and is still under

study to ma*e sure that it +ill #e sa,e and e2ective. ene thera$y is

currently only #eing tested ,or the treatment o, diseases that have no

other cures. It should #e noted that not all medical $rocedures that

introduce alterations to a $atient?s genetic ma*eu$ can #e considered

gene thera$y. Bone marro+ trans$lantation- and organ trans$lants ingeneral have #een ,ound to introduce ,oreign GA into $atients. ene

thera$y is de4ned #y the $recision o, the $rocedure and the intention o, 

direct thera$eutic e2ects. 

ene thera$y +as conce$tualied in 7F5- #y authors +ho urged

caution #e,ore commencing human gene thera$y studies.

 The 4rst attem$t- al#eit an unsuccess,ul one- at gene thera$y 'as +ell

as the 4rst case o, medical trans,er o, ,oreign genes into humans notcounting organ trans$lantation) +as $er,ormed #y Martin Cline on 76

 uly 7FD6. Cline claimed that one o, the genes in his $atients +as active

si months later- though he never $u#lished this data or had it

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veri4ed and even i, he is correct- it?s unli*ely it $roduced any signi4cant

#ene4cial e2ects treating #eta9thalassemia.

 The 4rst germ line gene thera$y consisted o, $roducing a genetically

engineered em#ryo in Octo#er 7FF:. The #a#y +as #orn on uly 57-

7FF and +as $roduced #y ta*ing a donor?s egg +ith healthymitochondria- removing its nuclear GA and 4lling it +ith the nuclear

GA o, the #iological mother 9 a $rocedure *no+n as cyto$lasmic

trans,er.

 This $rocedure +as re,erred to sensationally and some+hat inaccurately

in the media as a <three $arent #a#y<- though mtGA is not the $rimary

human genome and has little e2ect on an organism?s individual

characteristics #eyond $o+ering their cells.

ene thera$y is a +ay to 4 a genetic $ro#lem at its source. The

$olymers are either e$ressed as $roteins- inter,ere +ith $rotein

e$ression- or $ossi#ly correct genetic mutations.

 The most common ,orm uses GA that encodes a ,unctional-

thera$eutic gene to re$lace a mutated gene. The $olymer molecule is

$ac*aged +ithin a <vector<- +hich carries the molecule inside cells.

 The 4rst commercial gene thera$y- endicine- +as a$$roved in China in566L ,or the treatment o, certain cancers. In 5677 Geovasculgen +asregistered in @ussia as the 4rst9in9class gene9thera$y drug ,or treatmento, $eri$heral artery disease- including critical lim# ischemia. In5675 ly#era- a treatment ,or a rare inherited disorder- #ecame the 4rsttreatment to #e a$$roved ,or clinical use in either Euro$e or the UnitedStates a,ter its endorsement #y the Euro$ean Commission. 

A!A de2cienc-  is one ,orm o, SCI 'severe com#inedimmunode4ciency)- a disorder that a2ects the immune system. AA

de4ciency is very rare- #ut very dangerous- #ecause a mal,unctioningimmune system leaves the #ody o$en to in,ection ,rom #acteria andviruses.

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 The disease is caused #y amutation in a gene onchromosome 56.  A!Ade2cienc- is inherited inan aut,s,0al recessi1e0anner. The gene codes ,orthe enyme adenosinedeaminase 'AA). =ithoutthis enyme- the #ody isuna#le to #rea* do+n a toicsu#stance calleddeoyadenosine. The toin#uilds u$ and destroysin,ection94ghting immunecells called T and B

lym$hocytes. Because AAde4ciency a2ects theimmune system- $eo$le +ho have the disorder are more susce$ti#le toall *inds o, in,ections- $articularly those o, the s*in- res$iratory system-and gastrointestinal tract. They may also #e shorter than normal. Sadly-most #a#ies +ho are #orn +ith the disorder die +ithin a ,e+ months.

$reat0ents ,/ A!A !e2cienc- includes"

• #one marro+ trans$lant

  gene therap-

• AA enyme in PE vehicle

'n Septe0ber 8<4 899=4 the 2rst gene therap-  to com#at this

disease +as $er,ormed #y r. =illiam rench Anderson on a /,ur>-ear>

,ld girl4 Ashanti !eSil1a- at the Gational Institutes o, (ealth-

Bethesda- Maryland- U.S.A.

ConclusionBiotechnology is the ne+ +onder o, science. It is truly multidisci$linaryin nature and it encom$asses several disci$lines o, #asic sciences andengineering. The Science disci$lines ,rom +hich #iotechnology dra+sheavily are micro#iology- chemistry- #iochemistry- genetics- molecular#iology- immunology- cell and tissue culture and $hysiology. On the

engineering side it leans heavily on $rocess chemical and #iochemicalengineering since large scale cultivation o, microorganisms and cells-their do+nstream $rocessing are #ased on them. It c,0es t, us as agreat blessing...

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Biotechnology utilies the techni3ue called genetic engineering orrecom#inant GA technology +here a microorganism is isolated itsgenetic material is cut- mani$ulated- sealed- again inserted in anorganism and allo+ed to gro+ in a suita#le environment undercontrolled conditions to get the desired $roduct. It loo*s easy #ut is avery tedious jo# and it ta*es years ,or a research to achieve its goal.i*e every other thing- bi,techn,l,g- t,, has s,0e har0/uli0pacts"

7. enetic engineering is a very vital $art o, #iotechnology and thecost o, trans,erring genes ,rom one s$ecies to another is verye$ensive- +hich re3uires a huge amount o, ca$ital investment.$he c,st ,/ pr,ducing geneticall-> 0,di2ed plants andani0als are s7-> r,c7eting and the duration o, return are alsonot $redicta#le.

5. enetic engineering crosses #oundaries o, re$roduction #y

crossing genes o, s$ecies that are com$letely unrelated hencegiving rise to haardous results as +ell as also increasing the ris*o, harming multi$le s$ecies.

L. =hen genetic material ,rom certain viruses is used in the$roduction o, transgenic cro$s- there are chances that these virusgenes +ill com#ine +ith cro$ genes to $roduce more destructiveviruses. The consum$tion o, such cro$s is haardous to humanhealth and can cause several li,e9 threatening ailments. It can alsoresult in cancer- o,ten malignant as +ell.

H. Biotechnology also $oses a num#er o, environmental threats.

enetically modi4es cro$s o,ten in,ect monarch #utteries andother insect s$ecies.

 The a$$lications o, #iotechnology are so #road- and the advantages so

com$elling- that virtually every industry is using this technology.

evelo$ments are under+ay in areas as diverse as $harmaceuticals-

diagnostics- tetiles- a3uaculture- ,orestry- chemicals- household

$roducts- environmental cleanu$- ,ood $rocessing and ,orensics to name

a ,e+. Biotechnology is ena#ling these industries to ma*e ne+ or #etter

$roducts- o,ten +ith greater s$eed- e>ciency and Nei#ility.Bi,techn,l,g- 0ust c,ntinue t, be care/ull- regulated s, that

the 0a3i0u0 bene2ts are recei1ed 5ith the least ris7 .

Bi#liogra$hyhttp?@@en.5i7ipedia.,rg@bi,techn,l,g-http?@@en.5i7ipedia.,rg@insulin

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http?@@555.gene5atch.,rg@sub>:;:http?@@en.5i7ipedia.,rg@hu0ulinhttp?@@555.bi,techarticles.c,0@'thers>Article@Hu0an>

Insulin>and>#ec,0binant>!NA>$echn,l,g->D=.ht0lhttps?@@isaaa.,rg@res,urces@publicati,ns@p,c7et7@<@de/ault.

asphttp?@@555.sciencedirect.c,0@https?@@en.5i7ipedia.,rg@5i7i@GeneEtherap-https?@@en.5i7ipedia.,rg@5i7i@Aden,sineEdea0inaseEde2cie

nc-http?@@555.diabetes.c,.u7@insulin@ani0al>insulin.ht0lBi,l,g- te3tb,,7 )N.C.".#.$* Class 8;th

Contents

• Introduction

• (istory

•

Biotechnology in Agriculture• enetically Modi4ed Cro$s

• @GA Inter,erence '@GAi)

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• Bt toin

• Bt cotton

• Biotechnology in Medicine

• enetically engineered insulin'(umulin)

• ene thera$y

• Conclusion

• Bi#liogra$hy