Team:The University of Alberta/22 July 2008

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Latest revision as of 01:38, 23 July 2008

Mike added links to two protein complementation papers in the Project/Journals section of the Wiki. One of these uses a Vibrio transcription factor (ToxR) fragment and Vibrio promoter in E. coli, and the other uses B-galactosidase in E. coli. Can anyone figure out how to make the appropriate BioBricks to fuse these to the existing ER LBD BioBrick? We should be able to amplify the B-galactosidase fragments from E. coli, but we will need to synthesize the Vibrio fragments and promoter. We will also need to order a lacZ deletion strain of E. coli (listed in the paper) if we don't already have something like it.

Can anyone commit to designing these primers and synthetic constructs (ASAP?)

Did minipreps on O/N's of I0500 (new) col 1 and col 2. They are kept in Miniprep box 2 in freezer.

David digested BisdA and BisdB for insertion into 2 different vectors each:

I digested BisdA and BisdB with both (NgoMIV+Age1) and (XbaI+PstI) and concentrated all of the digests down to 30ul.

I digested the vector containing the TetR binding site and the RBS with NgoMIV and treated the digest with Antartic Phosphatase. I also concentrated this digest down to 30ul.

I digested J61003 with (SpeI+PstI) and treated the digest with Antartic Phosphatase. I also concentrated this digest down to 30ul.

The concentrated digests have been kept at -20C overnight and should be gel purified tomorrow, with the BisdA/B genes being purified and inserted into the digested vectors.

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-Mike added links to two protein complementation papers in the Project/Documents section of the Wiki.  One of these uses a Vibrio transcription factor (ToxR) fragment and Vibrio promoter in E. coli, and the other uses B-galactosidase in E. coli.  Can anyone figure out how to make the appropriate BioBricks to fuse these to the existing ER LBD BioBrick?  We should be able to amplify the B-galactosidase fragments from E. coli, but we will need to synthesize the Vibrio fragments and promoter.  We will also need to order a lacZ deletion strain of E. coli (listed in the paper) if we don't already have something like it.
+Mike added links to two protein complementation papers in the Project/Journals section of the Wiki.  One of these uses a Vibrio transcription factor (ToxR) fragment and Vibrio promoter in E. coli, and the other uses B-galactosidase in E. coli.  Can anyone figure out how to make the appropriate BioBricks to fuse these to the existing ER LBD BioBrick?  We should be able to amplify the B-galactosidase fragments from E. coli, but we will need to synthesize the Vibrio fragments and promoter.  We will also need to order a lacZ deletion strain of E. coli (listed in the paper) if we don't already have something like it.
+Can anyone commit to designing these primers and synthetic constructs (ASAP?)
+* Did minipreps on O/N's of I0500 (new) col 1 and col 2. They are kept in Miniprep box 2 in freezer.
+David digested BisdA and BisdB for insertion into 2 different vectors each:
+I digested BisdA and BisdB with both (NgoMIV+Age1) and (XbaI+PstI) and concentrated all of the digests down to 30ul.
+I digested the vector containing the TetR binding site and the RBS with NgoMIV and treated the digest with Antartic Phosphatase.  I also concentrated this digest down to 30ul.
+I digested J61003 with (SpeI+PstI) and treated the digest with Antartic Phosphatase.  I also concentrated this digest down to 30ul.
+The concentrated digests have been kept at -20C overnight and should be gel purified tomorrow, with the BisdA/B genes being purified and inserted into the digested vectors.