Some useful statistics:

Small genome (114.5 Mb/125 Mb total)

has been sequenced in the year 2000

(SequenceViewer, AGI).

Extensive genetic and physical maps

of all 5 chromosomes (MapViewer).

A rapid life cycle (about 5-8 weeks from

germination to mature seed).

Prolific seed production and easy

cultivation in restricted space.

Efficient transformation methods

utilizing Agrobacterium tumefaciens.

SIGnAL "T-DNA Express" Arabidopsis Gene Mapping Tool

Moving forward in reverse: genetic technologies to

enable genome-wide phenomic screens in Arabidopsis

Jose M. Alonso* and Joseph R. Ecker (2006) Nature

Work PlanScreening of Arabidopsis mutants on MS plates

Planting mutant lines on Soil

DNA isolation

Amplify the flanking sequences of each insertion

(Using gene specific Primers)

Homozygosity and Heterozygosity analysis for T-DNA

through PCR

Identification of interesting phenotype linked with

homozygous T-DNA line

Amplification of Gene from wild type

Cloning and Sequencing of gene

Construct designing into pBINAR expression vector

Transformation into heterozygous mutant plant

Complementation of mutant phenotype

Germination of the different plant lines on MS medium

The resistant plants of mutant 165B07

are germinated on MS having

sulfadiazine

The resistant plants of mutant 060B06 are

germinated on MS having sulfadiazine

Description of the Mutant

Phenotype of GABI mutant

Disrupted gene:

The disrupted gene was At5g16980

genomic sequence length: 1601 nucleotides

coding sequence length: 701 nucleotides.

T-DNA Sequence

AATCTGTGCGCGGGAATTATCGCATAAATTGGCTTTGGGATCCTCCC

TATGGGAGCAGAAGACTCAACGCGATAGAGATAGCGAAAAGAAAAA

CCCGAAGAGACACAATTTCTTCTCAGAGTCTTCAAGAACACCAAAT

TGTCATGTAGTTTCTACATCAATAAGAAGCATCTA

Gene AnnotationThe gene At5g16980 codes for a putative NADP-dependent

oxidoreductase like protein Protein propertiesLength : 311 amino acids

molecular weight : 34528.9 Da

Isoelectric point is 6.32

Disrupted gene

At4g29060 which has 4312 nucleotides for genomic DNA

sequences and 2862 nucleotides for coding sequences.

T-DNA Sequence

GGGTTCATACTCATTGCTGGTGTCTCTCCCCTGTGCTATTTTTATTTCGTTTT

CCCTCTCTTGGTGGATAGAGGAGATAAGAGTCTTCAAAGATTACTACAGGTGA

TTGTCTTTTTCTCTCTCTCCTGCGCCATCAAGACTCGAGCTTCAATATGGGCT

TACGGGCTTCATACTAAGGCCCATCAATAGCCTATATCGATGTGAAACTCCCC

CCCAGACATTACATAGTAGCAATGCCTCCTTTTCATGCTGGTCAATTTCTTAA

TAAATCAATAAGAGGCGTCGAACAAGACTTATTCAACATTTTATTACTAAGAGT

CTAAAAGACTTTAGTTTTTGACCCTCCAAGTCATACAGATAAAGCCTATTTAGA

TCTTTTTCATATTATAATCATATGGTTTTGACCAAAAAAAGAAAAACTTATGGT

AAGATCATTCATGAGAGGAAAAAAGAATTGCCAAACGATGA

TCTACCACTCTCCTCGTTTTCGCTTCGTGCTTATTTGTCAAACGCGAACATTTCA

CCGAAACTTAGCGACTTTTCTCGCTTTACTCTTTCTGGGAGCAATTTCTCAGAG

CATAGGCGAGCTTAAAGGTAGCTCCTTTTCTTAGCTTTGCTCAAATGGGTTTCT

TAGTTACTCTTCTTATGTTTCCGTGATAAGATGGTTCGAAATTTACTATGGGAT

CCTCCGTATAGTGAGGCGTTNTTACTAANNNNNNNNNNNNNNNNNNNNNN

NNNNNNNNNAAGAAAAAGAGTGGTCCTTTTCTTCATATGCTTTCAGATTAG

TCTGTGTTGTTGTCTGAGGAAAGCTTGTAATTTTGACAGTGTGTAATGGTTT

TCTATGTGTAGGTGACAGTTCTTAGCTATGGCTACGATTACACCTTCTTCTAT

TAGCAATGCCTGGCTCATTCCTGGAGCTTCTTTTACTGTAAAGAAGAGTGAT

TGTTCCATCAAATGTA

Gene AnnotationAt4g29060 codes for a putative Quinone oxidoreductase like protein which

functions in chain elongation during polypeptide synthesis at the ribosome

Protein Properties

length : 953 amino acids

Molecular weight 103782 Da

Isoelectric point : 4.95

localized in chloroplast and Ribosome

PCR amplification of small pale green plants with gene specific forward and GK-

LB primer.

Amplification and cloning of the At5g59650 gene from Col-0

Genevestigator database (https://www.genevestigator.ethz.ch)

3.7Kb

3.7Kb

Restriction digestion of the cloned At5g59650 and pGEMT

vector 3.7kb, with Sal-1 and Xba-1 restriction enzymes.