2. Why Would You Want To Insert New DNA into E. coli? Insulin Gene Extracted Plasmid

3. My Focus Insulin Gene Extracted Plasmid

4. A Closer Look at Taking Up New DNA

5. What Is Competency?

6. How does natural competency work?

7. How does ComK activate late competency genes? K-box promoter ComK Late Competency Gene

8. How are cells made competent? Natural Competency in B. subtilis –Controlled by ComK Induced Competency in E. coli K-boxes 1014 Both 48 genes Microarray 57 genes

9. Is natural competency feasible in E. coli?

10. K-Box to gene distance not consistent.

11. Building a Testable System AmpR ComK High Copy MWM ComK 1018 bp 506 bp 396 bp 344 bp 298 bp 600 bps

12. Building a Testable System KanR ComK Low Copy RBS AmpR pKBox High Copy GFP

13. Testing Fluorescence p < 0.02

14. Building a Testable System KanR ComK Low Copy RBS AmpR pKBox High Copy GFP RBS

15. RBS’s: Not Consistent

16. Promoters: Consistent

17. Building a Competent System KanR ComK Low Copy

18. Testing Competency ComK / KanPellet CellsRFP / AmpLB + Kan/Amp

19. Conclusions In E. coli:In B. subtilis: pKBox + ComK

20. 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