RRC ID 33453
著者 Koop AH, Hartley ME, Bourgeois S.
タイトル A low-copy-number vector utilizing beta-galactosidase for the analysis of gene control elements.
ジャーナル Gene
Abstract A low-copy-number vector, pFZY1, with the multiple restriction site linker of M13mp18 inserted upstream from a promoterless beta-galactosidase (beta Gal)-coding lacZ gene has been constructed to provide a convenient and accurate system to analyze regulatory elements in vivo. The plasmid contains the oriF replication origin without the par locus and is present in the cell in one to two copies per genome. It is retained in the host by the presence of ampicillin, and each inserted promoter yielded consistent values of beta Gal activity under all the conditions tested. A series of tetracycline resistance (TcR) promoter fragments and lac promoter fragments have been compared in pFZY1 and the high-copy-number pKO-vector series. The transcriptional activity measured for different fragments containing the same TcR promoter varied within a six-fold range among the several constructs tested. Regulation of the wild-type lac promoter and mutants in pFZY1 was similar to that observed for lac promoters in the chromosome while their regulation in pKO-1mp18 was significantly affected by the high copy number, as expected.
巻・号 52(2-3)
ページ 245-56
公開日 1987-1-1
DOI 10.1016/0378-1119(87)90051-5
PII 0378-1119(87)90051-5
PMID 3038688
MeSH Base Sequence DNA Restriction Enzymes DNA Transposable Elements* Escherichia coli / enzymology Escherichia coli / genetics* Galactosidases / genetics* Genes* Genes, Bacterial* Genetic Vectors* Mutation Plasmids Promoter Regions, Genetic* beta-Galactosidase / genetics*
IF 2.984
引用数 95
WOS 分野 GENETICS & HEREDITY
リソース情報
原核生物(大腸菌) pFZY1