The search continues…

This is Erin reporting the latest on the search the elusive temperature sensitive phenotype.

  I knew it was too good to last. I had a sneaking suspicions my project was being too cooperative. I know this sounds cynical but I usually never go this long without weird or no results.

I have finally hit the point in my project where trial and error has become my only option.

When the mutation combinations were electroporated into B. thailandensis, plated, and patched for screening, neither combination yielded Ts phenotype.

The revertant plasmid and Ts control were transformed back into E. coli patched out and will be isolated for sequencing early next week. However, there is some funny business going on with the Ts phenotype.

Also, six plasmids were sent for sequencing that were mutated with multi-site mutagenesis with the last point mutation to be tested. Two out of six are positive for a D to G mutation at site 145 in the rep gene. This is exciting for me because I have not been able to isolate a plasmid with this single point mutation yet. This point mutation will also be screened for Ts in B. thailandensis early next week.

 I wish I had more to post but for the time being I will just have to keep plugging along and see what the next new development is.

 Thanks for reading and as always, enjoy.

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