Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro

Messing, J, Gronenborn B, Müller-Hill B, Hans Hopschneider P.  1977.  

Journal:

Proceedings of the National Academy of Sciences of the United States of America

Volume Number:

74

Pages:

3642-6

Abstract:

A HindII restriction fragment comprising the Escherichia coli lac regulatory region and the genetic information for the alpha peptide of beta-galactosidase (beta-D-galactosidegalactohydrolase, EC. 3.2.1.23) has been inserted into 1 of the 10 Bsu I cleavage sites of M13 by blunt end ligation. A stable hybrid phage was isolated and identified by its ability to complement the lac alpha function. Further characterization of the hybrid phage includes retransformation studies, agarose gel electrophoresis, DNA-DNA hybridization, and heteroduplex mapping. The insertion point has been localized at 0.083 map unit on thewild-type circular map-i.e., within the intergenic region. The results prove that part of the intergenic region is nonessential and that the phage can be used as a cloning vehicle.

Notes:

0027-8424333444

Related External URL:

http://www.ncbi.nlm.nih.gov/sites/entrez?db=PubMed&cmd=retrieve&dopt=AbstractPlus&list_uids=333444file://localhost/Users/messing/Documents/Lett.&Memos/New%20Text/Current%20Papers/Papers/1977/Messing/MessingProc%20Natl%20Acad%20Sci%20USA%201977.pdfpapers:/
Citation:
Messing, J, Gronenborn B, Müller-Hill B, Hans Hopschneider P.  1977.  Filamentous coliphage M13 as a cloning vehicle: insertion of a HindII fragment of the lac regulatory region in M13 replicative form in vitro. Proceedings of the National Academy of Sciences of the United States of America. 74:3642-6.