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[抗灰霉素基因gsr的克隆及其在灰色链霉菌Kr.菌株中的功能研究]

[Cloning of grisin resistance gene gsr and the study of its functioning in Streptomyces griseus Kr. strain].

作者信息

Sezonov G V, Kriugel' G, Kudriashova E A, Kolibaba L G, Lomovskaia N D

出版信息

Antibiot Khimioter. 1990 Dec;35(12):14-7.

PMID:2127665
Abstract

S. griseus Kr. is a commercial strain producing grisin, an antibiotic of the streptothricin group used as a feed additive. It was shown earlier that genetic instability of the strain was very high which was evident from a high frequency of nonreverting Grn- Grns mutants. With densitographic analysis of chromosomal DNA electrophoregrams and DNA-DNA hybridization it was revealed that the molecular basis of the genetic instability of the S. griseus strain was deletion of a DNA fragment about 20 kb in size containing a grisin resistance gene. The resistance gene designated as gsr was cloned to S. lividans TK 64 within the plasmid vector pIJ699. The restriction map of a cloned DNA fragment with a gsr gene was constructed and its similarity to that of a nat gene resistant to norseothricin, another streptothricin was observed. Introduction of a gsr gene within the multicopy plasmid pIJ699 into S. griseus 212, a highly productive strain synthesiing the antibiotic, led to an increase in its resistance and productivity. Proceeding from the preliminary data on possible linkage of a gsr gene and grisin biosynthesis genes, it appeared possible to use the cloned gene as a molecular probe in cloning the biosynthesis genes.

摘要

灰色链霉菌Kr.是一种生产grisin的商业菌株,grisin是链丝菌素类抗生素,用作饲料添加剂。早期研究表明,该菌株的遗传不稳定性非常高,这从非回复性Grn-Grns突变体的高频率中可见一斑。通过对染色体DNA电泳图谱的光密度分析和DNA-DNA杂交,发现灰色链霉菌株遗传不稳定性的分子基础是缺失了一个约20 kb大小的包含grisin抗性基因的DNA片段。将指定为gsr的抗性基因克隆到质粒载体pIJ699中的变铅青链霉菌TK 64中。构建了带有gsr基因的克隆DNA片段的限制性图谱,并观察到其与对另一种链丝菌素诺尔丝菌素具有抗性的nat基因的限制性图谱相似。将多拷贝质粒pIJ699中的gsr基因导入高产抗生素合成菌株灰色链霉菌212中,导致其抗性和生产力增加。根据关于gsr基因与grisin生物合成基因可能存在连锁的初步数据,似乎可以将克隆基因用作克隆生物合成基因的分子探针。

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