GENE-CLUSTER FOR STREPTOMYCIN BIOSYNTHESIS IN STREPTOMYCES-GRISEUS - ANALYSIS OF A CENTRAL REGION INCLUDING THE MAJOR RESISTANCE GENE
GENE-CLUSTER FOR STREPTOMYCIN BIOSYNTHESIS IN STREPTOMYCES-GRISEUS - ANALYSIS OF A CENTRAL REGION INCLUDING THE MAJOR RESISTANCE GENE
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DOI：
10.1007/bf00330443
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发表时间：
1987-06-01
期刊：
影响因子：
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通讯作者：
PIEPERSBERG, W
中科院分区：
文献类型：
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作者：
DISTLER, J;BRAUN, C;PIEPERSBERG, W
对从灰色链霉菌中克隆的抗菌素链霉素生物合成基因的中心片段进行了开放阅读框、转录和翻译活性的分析。核苷酸序列显示两个有意义的开放阅读框ORF1和APH(6)，方向相反，起始密码子之间有885bp的间隔区。第一个是ORF1，编码能力为38 kDa。开放阅读框APH(6)是链霉素6-磷酸转移酶的主要抗性基因，它由307个氨基酸残基组成，大小为33 kDa。对纯化的APH(6)酶蛋白的前14个N端氨基酸残基进行了序列测定，结果与所提出的一级结构一致。讨论了ORF1推测的基因产物与体外翻译蛋白(表观分子量41 kDa)的可能同一性。灰链霉菌和产羟链霉菌的两个APH(6)基因的比较。Vogtli和Hutter 1987)揭示了编码区75%的核苷酸序列同源性和74%的多肽序列保守性。检测到了在其他抗生素和蛋白磷酸转移酶中高度保守的两个蛋白结构域。
A central segment of a cluster of biosynthetic genes for the antibiotic streptomycin cloned from Streptomyces griseus was analysed for open reading frames, as well as for transcriptional and translatonal activity. The nucleotide sequence revealed two significant open reading frames, ORF1 and APH(6), orientated in opposite directions and with a spacer of 885 bp between the start codons. The first, ORF1, had a coding capcity of 38 kDa. One open reading frame, APH(6), was identified as the major resistance gene coding for streptomycin 6-phosphotransferase, a protein of 307 amino acid residues and 33 kDa. Sequence determination of the first 14 N-terminal amino acid residues of the purified APH(6) enzyme protein was in agreement with the proposed primary structure. The possible identity of the presumed gene product of ORF1 with an in vitro translated protein (apparent molecular weight 41 kDa) is discussed. Comparison of the two APH(6) genes from S. griseus and the hydroxystreptomycin-producing S. glaucescens (cf. Vogtli and Hutter 1987) revealed 75% nucleotide sequence homology in the coding region and 74% conservation of the polypeptide sequence. Two protein domains which are highly conserved in other antibiotic and protein phosphotransferases were detected.