Kozlovskaia T M, Dishler A V, Bychko V V, Pumpen P P, Gren E Ia
Mol Biol (Mosk). 1981 Sep-Oct;15(5):1158-68.
A series of plasmids with tetracycline resistance genes (Tcr-operon) subjected to transcription from chloramphenicol acetyl transferase promoter (Cmr-promoter) have been constructed on the basis of plasmid pBR325, AprCmrTcr. For this purpose, a 0.8 Md fragment in pBR325 DNA bordered by unique EcoRI and HindIII restriction sites was cut out and structural genes of Tcr-operon were fused to the cat gene nucleotides corresponding to Cmr-promoter and first 72 amino acids of cat (alton, Vapnek, 1979; Marcoli et al., 1980). These plasmids with molecular weight amounting to 3 Md confer AprTcr phenotype to host cells. Tetracycline resistance can be eliminated completely by the deletion of a) Cmr-promoter; b) part of the first Tcr-operon gene.
基于质粒pBR325(AprCmrTcr)构建了一系列带有四环素抗性基因(Tcr操纵子)且受氯霉素乙酰转移酶启动子(Cmr启动子)转录调控的质粒。为此,切出了pBR325 DNA中一个由独特的EcoRI和HindIII限制性位点界定的0.8 Md片段,并将Tcr操纵子的结构基因与对应于Cmr启动子和cat基因前72个氨基酸的核苷酸融合(阿尔顿、瓦普内克,1979;马尔科利等人,1980)。这些分子量为3 Md的质粒赋予宿主细胞AprTcr表型。通过缺失以下部分可完全消除四环素抗性:a)Cmr启动子;b)第一个Tcr操纵子基因的一部分。