Gang L, Wei C, Yuqing T, Huarong T, Chater K F, Buttner M J
Institute of Microbiology, The Chinese Academy of Sciences, Beijing.
Chin J Biotechnol. 1999;15(4):195-202.
A 1.3 kb DNA fragment was cloned from a total DNA library of Streptomyces ansochromogenes using Southern hybridization. Nucleotide sequencing analysis indicated that the 1320 bp DNA fragment contained a complete open reading frame (ORF). In search of databases, the deduced product of ORF containing 213 amino acids is homologous to the serine protease of Caulobacter cresceatus, and a conserved serine-catalytic active site (GPSAG) exists. The gene was designated as sawD. The function of this gene was studied with the strategy of gene disruption, and the result showed that the sawD may be related to sporulation and especially to the spore septation in Streptomyces ansochromogenes. The preliminary result indicated that sawD mutant could produce abundant pigment in contrast with the wild type, it seems that sawD gene may be involved in pigment biosynthesis, and this gene is also dispensable for biosynthesis of nikkomycin in Streptomyces ansochromogenes.
使用Southern杂交技术从产色链霉菌的总DNA文库中克隆出一个1.3 kb的DNA片段。核苷酸测序分析表明,这个1320 bp的DNA片段包含一个完整的开放阅读框(ORF)。在数据库搜索中,包含213个氨基酸的ORF推导产物与新月柄杆菌的丝氨酸蛋白酶同源,并且存在一个保守的丝氨酸催化活性位点(GPSAG)。该基因被命名为sawD。采用基因破坏策略对该基因的功能进行了研究,结果表明sawD可能与产色链霉菌的孢子形成有关,特别是与孢子隔膜形成有关。初步结果表明,与野生型相比,sawD突变体能够产生丰富的色素,似乎sawD基因可能参与色素生物合成,并且该基因对于产色链霉菌中尼可霉素的生物合成也是非必需的。