Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
Download Inserting a Competency Regulatory Gene into E. coli
Document related concepts
Molecular cloning wikipedia , lookup
DNA vaccination wikipedia , lookup
Essential gene wikipedia , lookup
Cancer epigenetics wikipedia , lookup
Public health genomics wikipedia , lookup
Gene nomenclature wikipedia , lookup
Polycomb Group Proteins and Cancer wikipedia , lookup
Ridge (biology) wikipedia , lookup
Point mutation wikipedia , lookup
Genomic imprinting wikipedia , lookup
Oncogenomics wikipedia , lookup
Minimal genome wikipedia , lookup
Gene expression programming wikipedia , lookup
Gene therapy of the human retina wikipedia , lookup
Pathogenomics wikipedia , lookup
Gene therapy wikipedia , lookup
Genome evolution wikipedia , lookup
Gene desert wikipedia , lookup
Genetic engineering wikipedia , lookup
Epigenetics of human development wikipedia , lookup
Epigenetics of diabetes Type 2 wikipedia , lookup
Genomic library wikipedia , lookup
Vectors in gene therapy wikipedia , lookup
Biology and consumer behaviour wikipedia , lookup
Genome (book) wikipedia , lookup
Helitron (biology) wikipedia , lookup
Nutriepigenomics wikipedia , lookup
Therapeutic gene modulation wikipedia , lookup
Gene expression profiling wikipedia , lookup
Site-specific recombinase technology wikipedia , lookup
History of genetic engineering wikipedia , lookup
Designer baby wikipedia , lookup
Microevolution wikipedia , lookup
Artificial gene synthesis wikipedia , lookup
No-SCAR (Scarless Cas9 Assisted Recombineering) Genome Editing wikipedia , lookup
Transcript
Why Would You Want To Insert New DNA into E. coli? Insulin Gene Extracted Plasmid My Focus Insulin Gene Extracted Plasmid A Closer Look at Taking Up New DNA What Is Competency? How does natural competency work? How does ComK activate late competency genes? ComK K-box promoter Late Competency Gene How are cells made competent? • Natural Competency in B. subtilis – Controlled by ComK K-boxes 1014 Both Microarray 48 57 genes genes • Induced Competency in E. coli Is natural competency feasible in E. coli? Distance to Kbox in E. coli (bps) 22000 20000 18000 16000 14000 12000 10000 8000 6000 4000 2000 0 0 2000 4000 6000 8000 10000 12000 14000 16000 18000 20000 Distance to Kbox in B.subtilis (bps) 22000 24000 26000 Is natural competency feasible in E. coli? Distance to Kbox in E. coli (bps) 22000 20000 K-Box to gene distance not consistent. 18000 16000 14000 12000 10000 8000 6000 4000 2000 0 0 2000 4000 6000 8000 10000 12000 14000 16000 18000 20000 Distance to Kbox in B.subtilis (bps) 22000 24000 26000 Building a Testable System 600 bps ComK ComK MWM 1018 bp 506 bp 396 bp 344 bp 298 bp Building a Testable System RBS ComK pKBox GFP Testing Fluorescence 12 10 rlu/OD 8 6 p < 0.02 4 2 0 GFP pKBox:B0032:GFP pKBox:B0032:GFP +ComK Building a Testable System RBS ComK RBS pKBox GFP RBS’s: Not Consistent 210 180 150 rlu/OD 120 90 60 30 0 Promoters: Consistent 210 180 150 rlu/OD 120 90 60 30 0 Building a Competent System ComK Testing Competency ComK / Kan Pellet Cells RFP / Amp LB + Kan/Amp Conclusions In E. coli: pKBox + ComK In B. subtilis: Acknowledgments • • • • • • • Davidson Research Initiative National Science Foundation Howard Hughes Medical Institute James G. Martin Genomics Program Dr. Heyer, Dr. Campbell and Dr. Denham Fellow lab workers Friends and Family
