TY - JOUR
T1 - Use of M13mp phages to study gene regulation, structure and function
T2 - cloning and recombinational analysis of genes of the Salmonella typhimurium histidine operon
AU - Artz, Stanley
AU - Holzschu, Donald
AU - Blum, Paul
AU - Shand, Richard
N1 - Funding Information:
We ure especially grateful to Joachim Messing and Gisellt Heidecker for providing phagesM 13mp8a nd Ml3mp9 and for instruction in their use.W e thank Allan Campbella nd WilliamT imberlakef or helpful advice,L isa Couperf or expertt echnicala ssistance, Dan Riggsf or help with the photographyL, ewanna Archer for typing,a nd John Ingrah~ and David Pratt for commentso n the manuscriptT. his work was supportedb y LJSPHS Grant GM27307.
PY - 1983/12
Y1 - 1983/12
N2 - A restriction map was determined for a φ80λdhis transducing phage DNA carrying the Salmonella typhimurium histidine operon. DNA fragments containing the promoter/regulatory region and the first two structural genes of the histidine operon (hisOGD) were identified by their ability to direct regulated synthesis of histidinol dehydrogenase (product of hisD) in a coupled in vitro protein synthesizing system. A 3.1-kb SalI-EcoRI restriction fragment containing the hisOGD region, was subcloned into phage M13mp8 and M13mp9 RF DNAs. Methods are described for shuttling mutant and wild-type bacterial DNA sequences between the M13mp:: his phage and host bacterial genomes. Of novel importance is the use of the phage M13 gene II amber mutation to obtain integration of the M13mp:: his phage genome into the homologous his region of the bacterial chromosome following transduction of recipients lacking an amber suppressor. This method can be used to facilitate allele replacement with genes carried on M13 transducing phages.
AB - A restriction map was determined for a φ80λdhis transducing phage DNA carrying the Salmonella typhimurium histidine operon. DNA fragments containing the promoter/regulatory region and the first two structural genes of the histidine operon (hisOGD) were identified by their ability to direct regulated synthesis of histidinol dehydrogenase (product of hisD) in a coupled in vitro protein synthesizing system. A 3.1-kb SalI-EcoRI restriction fragment containing the hisOGD region, was subcloned into phage M13mp8 and M13mp9 RF DNAs. Methods are described for shuttling mutant and wild-type bacterial DNA sequences between the M13mp:: his phage and host bacterial genomes. Of novel importance is the use of the phage M13 gene II amber mutation to obtain integration of the M13mp:: his phage genome into the homologous his region of the bacterial chromosome following transduction of recipients lacking an amber suppressor. This method can be used to facilitate allele replacement with genes carried on M13 transducing phages.
KW - Restriction mapping
KW - S1 nuclease
KW - in vitro protein synthesis
KW - shuttling DNA sequences
KW - site-specific mutagenesis
KW - specialized transduction
UR - http://www.scopus.com/inward/record.url?scp=0021018199&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0021018199&partnerID=8YFLogxK
U2 - 10.1016/0378-1119(83)90184-1
DO - 10.1016/0378-1119(83)90184-1
M3 - Article
C2 - 6323256
AN - SCOPUS:0021018199
VL - 26
SP - 147
EP - 158
JO - Gene
JF - Gene
SN - 0378-1119
IS - 2-3
ER -