Team:Illinois/SgrS

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SgrS Target-GFP Fusion

People in Group: Francis Lee, Harsh Shah, Dave Korenchan

Purpose: We want to fuse the SgrS target sequence from the PtsG gene to the GFP gene on a low-copy plasmid and transform this into E. coli cells along with a high-copy plasmid carrying the SgrS gene. We expect to see translational repression of GFP by the small RNA SgrS.

Protocol(s) Used: (links to protocols page)

Recipe(s) Used:

Primers Used:

June 15

PCRs of sRNA genes and target sequences were performed using E. coli extracted chromosomal DNA and primers.

June 16

We ran a gel on our PCR reactions, and the PCRs were successful! The SgrS gene falls at about the 300 bp region on our gel, which agrees with our data for the length of the SgrS gene segment. The PtsG target sequence is smaller than the SgrS gene on the gel, which also agrees with our data.

UI09Gel1.jpg