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Investigating Genetic Information Transfer via Bacterial Conjugation and Riboregulation

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This study focuses on the transfer of genetic information through bacterial conjugation, specifically utilizing F and RP4 plasmids. The F+ plasmid is crucial for this process, and the study aims to control transcription via riboregulation. The importance of pilus formation and the characteristics of conjugative plasmids are analyzed, with F and RP4 selected for their varying pili lengths and availability. Additionally, we explore gene knockouts essential for conjugation, specifically traJ on the F plasmid and potential new targets like trbC and traG.

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Investigating Genetic Information Transfer via Bacterial Conjugation and Riboregulation

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Presentation Transcript

  1. Implementation Goal: To transfer genetic information from one bacterial cell to another Means: Bacterial Conjugation Goal: To specifically control who can read the message Means: Riboregulation

  2. F F F Bacterial Conjugation • The F+ plasmid is necessary for bacterial conjugation F Pilus Formation F+ F- F+

  3. Why we used F and RP4 Plasmids • F and RP4 exhibit differing pili lengths, biasing the order in which F and RP4 will conjugate • F and RP4 do no conjugate with themselves • F and RP4 are among the most studied and well-characterized conjugative plasmids • F and RP4 plasmids are readily available

  4. Conjugative Plasmid Facts • Conjugative plasmids are very large, from 60k – 100k basepairs long • The Origin of Transfer (OriT) is where all DNA transfer begins during conjugation • Knocking out, by recombination, TraJ on the F plasmid, proved ineffective in stopping conjugation

  5. General Problems • All genes necessary, but to what degree? • traJ construct conjugates • Need to find new genes to knockout

  6. New Knockouts: What are we looking for? • Gene essential to conjugation that: • Will decrease conjugation by 7 orders of magnitude • Can be complemented in trans to restore conjugation

  7. New knockouts F plasmid  trbC gene R plasmid  traG

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