HELP! - cloning problem
ku at helix.nih.gov
Thu Nov 5 10:07:28 EST 1998
Perhaps you should be looking at the blue colonies rather than the white
ones. If your fragment is precisely the right size and does not contain
a termination codon you could be making a fusion protein with lacZ.
Alternatively if your fragment contains a sequence that can function as
a promoter/RBS you would still get lacZ activity. The white colonies
would then deletion derivatives of the full length clones.
> Thank you for the following comment. Yes, I have successfully cloned
> the whole coding region of the gene in a T-vector. It went just fine.
> I also cloned the gene from a different bacterial species, which is
> pretty similar to what I am going to clone, successfully into pBAD.
> So my guess is it may not be at least very toxic...?
Senior Staff Fellow
Section on Genomic Structure and Function
Laboratory of Molecular and Cellular Biology,
National Institutes of Diabetes and Kidney Diseases
Building 8, Room 202
8 Center Dr MSC 0830
National Institutes of Health
Bethesda, MD 20892-0830
email: ku at helix.nih.gov
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