Nie L P, Tian Y Q, Tan H R
Institute of Microbiology, The Chinese Academy of Sciences, Beijing 100080, China.
Yi Chuan Xue Bao. 2001;28(2):188-95.
A 7.5 kb DNA fragment was identified from the cosmid library of Streptomyces ansochromogenesus using a partial DNA fragment involved in the nikkomycin biosynthesis as a probe. It was cloned into pBluescript M13- and the resulting recombinant plasmid was designated as pNL2200. The nucleotide sequence of 2.3 kb Sal I -BamH I DNA fragment was determined and analysed. The result indicated that the fragment contains one complete open reading frame (ORF); The start codon is GTG at 271 position and the stop codon is TGA at 1,954 position. It contains 1,686 base pairs and encodes a protein with 561 amino acids. The deduced product has 44% identity with the ligase belong to the superfamily of adenylate-forming enzyme. The experiment of gene disruption showed that the sanJ gene was closely related to nikkomycins biosynthesis of Streptomyces ansochromogenes. It was designated as sanJ.
以参与多氧霉素生物合成的部分DNA片段为探针,从产色链霉菌的黏粒文库中鉴定出一个7.5 kb的DNA片段。将其克隆到pBluescript M13 - 中,所得重组质粒命名为pNL2200。测定并分析了2.3 kb Sal I - BamH I DNA片段的核苷酸序列。结果表明,该片段包含一个完整的开放阅读框(ORF);起始密码子位于第271位,为GTG,终止密码子位于第1954位,为TGA。它含有1686个碱基对,编码一个由561个氨基酸组成的蛋白质。推导的产物与属于腺苷酸形成酶超家族的连接酶有44%的同源性。基因破坏实验表明,sanJ基因与产色链霉菌的多氧霉素生物合成密切相关。将其命名为sanJ。