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M13 bacteriophage and pUC plasmids containing DNA inserts but still capable of beta-galactosidase alpha-complementation
Authors:T J Close  J L Christmann  R L Rodriguez
Institution:Department of Genetics, University of California, Davis, CA 95616 U.S.A. Tel. (916) 752-3263
Abstract:A DNA fragment encoding the transposon Tn9 chloramphenicol acetyltransferase gene (cat) was inserted into M13 phage and pUC plasmid cloning vehicles. When the cat gene was inserted in the same orientation as the lacZ gene, two new polypeptides were produced. One polypeptide possessed chloramphenicol acetyltransferase activity, while the other expressed beta-galactosidase alpha-donor activity. Both new polypeptides were translated from a hybrid messenger RNA initiating from the lac promoter. These observations may help explain why not all inserts produce white plaques.
Keywords:Insertional inactivation  blue plaques  chloramphenicol acetyltransferase  β-gal  β-galactosidase  bp  base pairs  CAT  chloramphenicol acetyltransferase  glc  glucose  IPTG  ssDNA  single-stranded DNA  X-gal  [ ]  indicates plasmid carrier state
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