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灰色链霉菌中灰霉素聚酮合酶基因簇的克隆、测序及分析

Cloning, sequencing, and analysis of the griseusin polyketide synthase gene cluster from Streptomyces griseus.

作者信息

Yu T W, Bibb M J, Revill W P, Hopwood D A

机构信息

John Innes Institute, John Innes Centre, Norwich, United Kingdom.

出版信息

J Bacteriol. 1994 May;176(9):2627-34. doi: 10.1128/jb.176.9.2627-2634.1994.

DOI:10.1128/jb.176.9.2627-2634.1994
PMID:8169211
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC205401/
Abstract

A fragment of DNA was cloned from the Streptomyces griseus K-63 genome by using genes (act) for the actinorhodin polyketide synthase (PKS) of Streptomyces coelicolor as a probe. Sequencing of a 5.4-kb segment of the cloned DNA revealed a set of five gris open reading frames (ORFs), corresponding to the act PKS genes, in the following order: ORF1 for a ketosynthase, ORF2 for a chain length-determining factor, ORF3 for an acyl carrier protein, ORF5 for a ketoreductase, and ORF4 for a cyclase-dehydrase. Replacement of the gris genes with a marker gene in the S. griseus genome by using a single-stranded suicide vector propagated in Escherichia coli resulted in loss of the ability to produce griseusins A and B, showing that the five gris genes do indeed encode the type II griseusin PKS. These genes, encoding a PKS that is programmed differently from those for other aromatic PKSs so far available, will provide further valuable material for analysis of the programming mechanism by the construction and analysis of strains carrying hybrid PKS.

摘要

以天蓝色链霉菌的放线紫红素聚酮合酶（PKS）基因（act）为探针，从灰色链霉菌K-63基因组中克隆出一段DNA片段。对克隆的DNA的一个5.4kb片段进行测序，发现了一组五个gris开放阅读框（ORF），它们对应于act PKS基因，顺序如下：酮合酶的ORF1、链长决定因子的ORF2、酰基载体蛋白的ORF3、酮还原酶的ORF5和环化酶-脱水酶的ORF4。通过使用在大肠杆菌中繁殖的单链自杀载体，将灰色链霉菌基因组中的gris基因替换为标记基因，导致产生灰霉素A和B的能力丧失，这表明这五个gris基因确实编码II型灰霉素PKS。这些基因编码的PKS与目前已知的其他芳香族PKS的编程方式不同，通过构建和分析携带杂交PKS的菌株，将为分析编程机制提供进一步有价值的数据。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e1b/205401/9e7e31874458/jbacter00027-0164-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e1b/205401/2be721196460/jbacter00027-0164-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e1b/205401/9e7e31874458/jbacter00027-0164-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e1b/205401/2be721196460/jbacter00027-0164-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2e1b/205401/9e7e31874458/jbacter00027-0164-b.jpg

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