CLONING AND EXPRESSION OF A TYLOSIN RESISTANCE GENE FROM A TYLOSIN-PRODUCING STRAIN OF STREPTOMYCES-FRADIAE

被引：52

作者：

BIRMINGHAM, VA ^{[1
]}

COX, KL ^{[1
]}

LARSON, JL ^{[1
]}

FISHMAN, SE ^{[1
]}

HERSHBERGER, CL ^{[1
]}

SENO, ET ^{[1
]}

机构：

[1] ELI LILLY & CO, DIV BIOCHEM DEV, LILLY CORP CTR, INDIANAPOLIS, IN 46285 USA

来源：

MOLECULAR AND GENERAL GENETICS | 1986年 / 204卷 / 03期

关键词：

D O I：

10.1007/BF00331036

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A gene conferring high-level resistance to tylosin in Streptomyces lividans and Streptomyces griseofuscus was cloned from a tylosin-producing strain of Streptomyces fradiae. The tylosin-resistance (Tylr) gene (tlrA) was isolated on five overlapping DNA fragments which contained a common 2.6 Kb KpnI fragment. The KpnI fragment contained all of the information required for the expression of the Tylr phenotype in S. lividans and S. griseofuscus. Southern hybridization indicated that the sequence conferring tylosin resistance was present on the same 5 kb SalI frament in genomic DNA from S. fradiae and several tylosin-sensitive (Tyls) mutants. The cloned tlrA gene failed to restore tylosin resistance in two Tyls mutants derived by protoplast formation and regeneration, and it restored partial resistance in a Tyls mutant obtained by N-methyl-N''-nitro-N-nitrosoguanidine (MNNG) mutagenesis. The tlrA gene conferred resistance to tylosin, carbomycin, niddamycin, vernamycin-B and, to some degree, lincomycin in S. griseofuscus, but it had no effect on sensitivity to streptomycin or spectinomycin, suggesting that the cloned gene is an MLS (macrolide, lincosamide, streptogramin-B)-resistance gene. Twenty-eight kb of S. fradiae DNA surrounding the tlrA gene was isolated from a genomic library in bacteriophage .lambda. Charon 4. Introduction of these DNA sequence into S. fradiae mutants blocked at different steps in tylosin biosynthesis failed to restore tylosin production, suggesting that the cloned Tylr gene is not closely linked to tylosin biosynthetic genes.

引用

页码：532 / 539

页数：8

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