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Structural and Functional Analysis of Pleiotropic Gene involved in Morphological and Metabolic Differentiation in Streptomyces

链霉菌形态和代谢分化相关多效基因的结构和功能分析

基本信息

批准号：
06660115
负责人：
OGATA Seiya
金额：
$ 1.34万
依托单位：
Kyushu University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

项目摘要

This report describes the peculiarity of a strain BalA1 derived from Thiostrepton-producing streptomyces azureus PK100C and the structure and function of its kpnI-genomic fragment of 2.5kb cloned in a vector pMCP5.The 2.5kb DNA-inserting plasmid was named as pSAB931. The pSAB931-harboring BalA1 showed the following specific characters on the crossfeeding solid culture : It was impossble to form the aerial mycelia and spores by itself, but parent PK100C and its wild-type strain PKO, another Thiostrepton-producing S.laurentii and some other streptomycetes stimulated the formation of those of BalA1 ; it inhibited the growth of strains PK100C and PKO, S.laurentii and some other streptomycetes ; it induced the production of a red color antibiotic (Undecylprodigiosin) of S.coelicolor at early stage of growth.The 2.5kb fragment of pSAB931 was used to probe Southern blots in order to detect the presence of the homologus sequence in diverse bacterial genome. Blotting analysis allowed us to detect the hybridized signals on all twenty streptomyces species tested (strains PK100C and PKO, S.laurentii, S.coelicolor etc). This result shows that this sequence presents in the genome of many Streptomyces species and it is an important gene.The DNA fragment was sequenced. It contained 2498bp. The frame analysis revealed two open reding frames, ORF843 and ORF489. They had 843bp and 489bp, respectively. The amino acid sequences of their deduced products were compared with those registered in the databases. The putative ORF843 product was shown a high similarity with phosphoserine phophatase (psp product) of Mycobacterium leprae. It was also shown that the putative ORF489 product was similar to a seputm placement determinants (minD product) of Bacillus subtilis.These results suggest that two ORFs are the pleiotropic genes involved in the morphological and metabolic differentiation.

本文报道了从硫链菌素产生菌蓝色链霉菌PK 100C中分离得到的BalA 1菌株的特性及其克隆到载体pMCP 5中的2.5kb kpn Ⅰ基因组片段的结构和功能。BalA 1在异交固体培养中表现出以下特性：BalA 1本身不能形成气生菌丝和孢子，但亲本PK 100C及其野生型菌株PKO、另一种产硫链霉菌罗伦链霉菌和其他一些链霉菌能促进BalA 1气生菌丝和孢子的形成;对PK 100C、PKO、罗伦链霉菌等链霉菌的生长有抑制作用;它诱导了一种红色抗生素的产生用pSAB 931的2.5kb片段进行Southern杂交，以检测其在不同细菌基因组中是否存在同源序列。印迹分析使我们能够检测到所有20种测试的链霉菌物种（菌株PK 100C和PKO、罗伦链霉菌、天蓝色链霉菌等）上的杂交信号。这一结果表明，该序列存在于许多链霉菌的基因组中，是一个重要的基因。它包含2498 bp。框架分析发现了两个开放的阅读框架，ORF 843和ORF 489。它们的长度分别为843 bp和489 bp。将推导产物的氨基酸序列与数据库中的氨基酸序列进行比较。推测的ORF 843产物显示与麻风分枝杆菌的磷酸丝氨酸磷酸酶（psp产物）高度相似。推测的ORF 489产物与枯草芽孢杆菌的一个seputm位置决定因子（minD产物）相似，表明这两个ORF是参与形态和代谢分化的多效性基因。