RRC ID 33470
著者 Casadaban MJ, Chou J, Cohen SN.
タイトル In vitro gene fusions that join an enzymatically active beta-galactosidase segment to amino-terminal fragments of exogenous proteins: Escherichia coli plasmid vectors for the detection and cloning of translational initiation signals.
ジャーナル J Bacteriol
Abstract We report the construction and use of a series of plasmid vectors suitable for the detection and cloning of translational control signals and 5' coding sequences of exogenously derived genes. In these plasmids, the first eight codons of the amino-terminal end of the lactose operon beta-galactosidase gene, lacZ, were removed, and unique BamHI, EcoRI, and SmaI (XmaI) endonuclease cleavage sites were incorporated adjacent to the eighth codon of lacZ. Introduction of deoxyribonucleic acid fragments containing appropriate regulatory signals and 5' coding sequences into such lac fusion plasmids led to the production of hybrid proteins consisting of the carboxyl-terminal segment of a beta-galactosidase remnant plus a peptide fragment that contained the amino-terminal amino acids encoded by the exogenous deoxyribonucleic acid sequence. These hybrid peptides retained beta-galactosidase enzymatic activity and yielded a Lac+ phenotype. Such hybrid proteins are useful for purifying peptide sequences encoded by exogenous deoxyribonucleic acid fragments and for studies relating the structure and function of specific peptide segments.
巻・号 143(2)
ページ 971-80
公開日 1980-8-1
DOI 10.1128/jb.143.2.971-980.1980
PMID 6162838
PMC PMC294402
MeSH Base Sequence DNA, Recombinant* Escherichia coli / genetics Galactosidases / genetics* Lac Operon Plasmids Protein Biosynthesis* RNA, Bacterial / genetics RNA, Messenger / genetics* Transcription, Genetic beta-Galactosidase / genetics* beta-Galactosidase / metabolism
IF 3.006
引用数 1011
WOS 分野 MICROBIOLOGY
リソース情報
原核生物(大腸菌) pMC931