Li Wenli, Tan Huarong
Department of Molecular Microbiology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, PR China.
Curr Microbiol. 2003 Jun;46(6):403-7. doi: 10.1007/s00284-002-3892-5.
A 6.3-kb BamHI- BglII DNA fragment was cloned from cos20 by using chromosome walking strategy. It was partially sequenced with the result that there is a possible ORF of 1272 nucleotides. The ORF designated sanV was deposited in GenBank under accession no. AF469955. Database search indicated that the deduced protein of sanV shows 28% identity and 44% similarity over 405 amino acid residues to the large component (E) of glutamate mutases from Clostridium cochlearium. Gene disruption was performed to study the function of sanV. It was found that sanV disruptants exhibited much poorer inhibition to the test strain than the wild-type S. ansochromogenes 7100. Furthermore, HPLC analysis indicated that the sanV disruptants almost did not produce nikkomycins X and Z, whereas they accumulated new nikkomycins O(x) and O(z), which revealed that sanV is an important gene involved in the biosynthesis of the peptidyl moiety of nikkomycins.
采用染色体步移策略从cos20中克隆出一个6.3 kb的BamHI - BglII DNA片段。对其进行了部分测序,结果显示存在一个1272个核苷酸的可能开放阅读框(ORF)。该命名为sanV的ORF已以登录号AF469955存入GenBank。数据库搜索表明,sanV推导的蛋白质在405个氨基酸残基上与来自耳蜗梭菌的谷氨酸变位酶的大组分(E)具有28%的同一性和44%的相似性。进行基因破坏以研究sanV的功能。发现sanV破坏株对测试菌株的抑制作用比野生型产色链霉菌7100差得多。此外,高效液相色谱分析表明,sanV破坏株几乎不产生多氧霉素X和Z,而它们积累了新的多氧霉素O(x)和O(z),这表明sanV是参与多氧霉素肽基部分生物合成的一个重要基因。