Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Is My Gene important for seed development in plants?
Document related concepts
Genetically modified crops wikipedia , lookup
Cell-free fetal DNA wikipedia , lookup
No-SCAR (Scarless Cas9 Assisted Recombineering) Genome Editing wikipedia , lookup
Epigenetics in learning and memory wikipedia , lookup
Genome evolution wikipedia , lookup
Copy-number variation wikipedia , lookup
Genome (book) wikipedia , lookup
Point mutation wikipedia , lookup
Genetic engineering wikipedia , lookup
Saethre–Chotzen syndrome wikipedia , lookup
Gene expression profiling wikipedia , lookup
Nutriepigenomics wikipedia , lookup
The Selfish Gene wikipedia , lookup
Genome editing wikipedia , lookup
Gene therapy of the human retina wikipedia , lookup
Gene expression programming wikipedia , lookup
Vectors in gene therapy wikipedia , lookup
Neuronal ceroid lipofuscinosis wikipedia , lookup
Gene therapy wikipedia , lookup
History of genetic engineering wikipedia , lookup
Gene desert wikipedia , lookup
Epigenetics of diabetes Type 2 wikipedia , lookup
Site-specific recombinase technology wikipedia , lookup
Gene nomenclature wikipedia , lookup
Helitron (biology) wikipedia , lookup
Microevolution wikipedia , lookup
Therapeutic gene modulation wikipedia , lookup
Transcript
Is My Gene Important for Seed Development in Plants?? Gene: AT3G53370 Jonathan Milgrom Spring 2004 What Does My Gene Do? Transcription Factor* • Repressor of Spinach gene rps1 • Active in roots • Highly conserved in plant kingdom Transcription factor *Dao-Xiu Zhou, Cordelia Bisanz Seyer Knockout the gene and find out! • Engineer agrobacteria to insert tDNA • Grow mutant seeds • Genotype tDNA AT3G53370 Is This Function Integral to Seed Development ? Madison & Salk Provide the Knockouts Madison SP #4 Salk Exon SP #1 5’ 3’ AT3G53370 Knockout Analysis Madison project began with… Superpool PCR products amplified with RV (gene specific) and tdna primers 4/15/04 Superpool tDNA line/ Mutant Seed Plant Genotype Sp #4 (small length suggests insertion) Salk project began with… Mutant seeds Madison Results Identified Madison DNA Pool tDNA S P # 4 Madison PCR products amplified with RV (gene specific) & tdna primers 5/18/04 5’ SP #4 DNA pool #6 AT3G53370 RV 33’ SALK Results 145678 W C 9t W 1456789t C 7 Plants Genotyped… Plant # genotype 1 Homozygous tDNA Heterozygous Wild Type 4 5 6 7 8 9 * * * * * * * PCR products of various SALK lines amplified with Tubulin • Homozygous tDNA exist • Knockout is not embryo lethal PCR products of various SALK lines amplified with Does My Gene Play Some Role in Seed Development? Gene activity/mRNA accumulation patterns Scarlet Runner Bean equivalent Arabidopsis Gene Chip Activity AT3G53370 5000 RT-PCR of 6 organs Signal Detection Strength 4500 4000 3500 3000 2500 Wild Type 2000 Lec1 mutant 1500 Mutant 1000 500 Organ 14 day old embryo Wild Type Floral Buds Wild Type Stem Wild Type Leaves Wild Type PostMaturation Wild Type MidMaturation Wild Type Ovules 0 Next…? • Study rsp1 spinach gene equivalent in Arabidopsis • Study lec1 gene • Identify Madison tDNA line and Genotype
