Cloning Vectors for Eukaryotes

Introduction to Recombinant Genetics- Biology 350Index • Syllabus • CourseInfo LogIn • References • Assignment • Next

CLONING VECTORS FOR EUKARYOTES

Vectors For Yeast

The 2 µm plasmid is a naturally occuring 6 kb episomal plasmid found in yeast.

Selection is based on nutrition instead of drugs. Yeast cells that are auxotrophic

(unable to synthesize an essential component, like an amino acid) can only grow when

nutritionally supplemented, or when the missing nutritional function is provided for by

genes contained on a plasmid or extra chromosome.

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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Many eukaryotic vectors contain both eukaryotic and prokaryotic signals and can

be replicated in both types of cells. These are called shuttle vectors.

Often, much of the cloning is done in prokaryotic systems and then the recombinant

vectors are grown in eukaryotic cells.

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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Why not just express all genes in prokaryotes? Why transfer to eukaryotes for

expression?

YEp Yeast episomal plasmid

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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YIp - Yeast integrative plasmid

YRps - Yeast replicative plasmid

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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YACs - Yeast artificial chromosome (600-1400 kb capacity)

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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CLONING VECTORS FOR HIGHER PLANTS

Agrobacterium Tumefaciens

The system was derived from the bacteria that causes crown gall disease in plants.

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During infection, the Ti plasmid is integrated into the plant chromosomal DNA.

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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The functions of the Ti plasmid have been divided between two plasmid-like vectors

which are co transformed and operate as a binary vector system.

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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Transformed plants can be recovered.

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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A binary vector

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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Direct gene transfer randomly integrates supercoiled DNA into cells (no homology

required).

CLONING VECTORS FOR ANIMALS

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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Cloning Into Insect Cells

Drosophilia cells and the P vector

Baculovirus

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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Cloning In Mammals

Purposes for cloning into animals

Gene knockout

Production of recombinant proteins

Gene therapy

Vectors are often derived from viral components

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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© 2005 by CA Rinehart Index • Syllabus • CourseInfo LogIn • References • Assignment • Next

This material is intended for use only by WKU students registered for Biology 350. Other uses prohibited.

Cloning Vectors for Eukaryotes http://bioweb.wku.edu/courses/biol350/cloningvecteuk9/review.html

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