HELP! - cloning problem

Karen Usdin ku at helix.nih.gov
Thu Nov 5 10:07:28 EST 1998


Mari

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...?
> 
>      Mari




~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Karen Usdin
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

Tel: 301-496-2189
Fax: 301-402-0053
email: ku at helix.nih.gov
http://www.niddk.nih.gov/intram/people/kusdin.htm
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~



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