pMC1871 (lacZ) needed
pnorton at lac.jci.tju.edu
Wed Sep 18 13:21:18 EST 1996
In article <32385261.1E at pen.gulbenkian.pt>, jcosta at pen.gulbenkian.pt wrote:
> I would be very interested in getting plasmid pMC1871 (Shapira et al.,
> 1983), a plasmid with BamHI-excisable intact lacZ gene.
> Can someone make it available or indicate a commercial source?
> Thanks in advance,
> ***** JOAO VASCONCELOS COSTA, MD, PhD
> ***** Instituto Gulbenkian de Ciencia - Oeiras, Portugal
> ***** mailto:jcosta at pen.gulbenkian.pt
> ***** http://www.pen.gulbenkian.pt/v2/jvc.html
Pharmacia sells the plasmid, but it is NOT an intact lacZ gene, the
amino terminal ca. 7 amino acids are missing. Thus, you need to tack on at
least an initiator codon, best if it is within a context favorable for
translation in your organism of interest.
We have taken the pMC1871 lacZ cassette and added an amino terminus
that is fairly well optimized for expression in mammalian cells (see
Biochem. Mol. Biol. Int. 36:483-490, 1995). The entire coding sequence can
be excised with a number of enzymes, such as HindIII or SalI, but you
would need to do a partial digest with BamHI.
Feel free to email me if you need more information.
Pamela A. Norton, Ph.D. Assistant Professor of Medicine
Thomas Jefferson University
Philadelphia, PA 19107 p_norton at lac.jci.tju.edu
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