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[从乳酸链球菌111工业菌株中克隆β-半乳糖苷酶基因并在大肠杆菌细胞中表达,以及该基因向嗜热链球菌细胞的接合转移]

[Cloning the gene of beta-galactosidase from the industrial strain of Streptococcus lactis 111 in E. coli cells and conjugated transfer of this gene to Streptococcus thermophilus cells].

作者信息

Molotov S V, Duzhiĭ D E, Danilevich V N, Sukhodolets V V

出版信息

Mol Gen Mikrobiol Virusol. 1991 Apr(4):3-7.

PMID:1906985
Abstract

The ability of the industrial strains of Streptococcus lactis to synthesize the enzyme beta-galactosidase was studied. Five strains among sixteen were found to produce high levels of the enzyme. The beta-galactosidase gene in the most active strain Streptococcus lactis 111 was shown to be located on the 50 kb conjugative plasmid. The plasmid was transferred by conjugation into Streptococcus thermophilus cells and subsequently the gene for beta-galactosidase was studied in transconjugants. The beta-galactosidase gene from Streptococcus lactis 111 was subcloned in Escherichia coli cells on the plasmid pBR322. The gene was localized on the 4.8 kb BgIII fragment of DNA. Following the restriction of DNA by the Sau3A the gene was subcloned on the birepliconed plasmid vector pCB20 capable of replication in the Gram-negative as well as Gram-positive microorganisms. The recombinant derivatives of pCB20 were isolated that carry the beta-galactosidase gene on the DNA fragments of different size.

摘要

对乳酸链球菌工业菌株合成β-半乳糖苷酶的能力进行了研究。在16株菌株中,发现有5株能产生高水平的该酶。活性最强的乳酸链球菌111中的β-半乳糖苷酶基因位于50 kb的接合质粒上。该质粒通过接合转移到嗜热链球菌细胞中,随后在接合子中研究β-半乳糖苷酶基因。来自乳酸链球菌111的β-半乳糖苷酶基因在大肠杆菌细胞中被亚克隆到质粒pBR322上。该基因定位在4.8 kb的BgIII DNA片段上。用Sau3A对DNA进行酶切后,该基因被亚克隆到能够在革兰氏阴性和革兰氏阳性微生物中复制的双复制子质粒载体pCB20上。分离得到了pCB20的重组衍生物,它们在不同大小的DNA片段上携带β-半乳糖苷酶基因。

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