激光生物学报, 2011, 20 (3): 367, 网络出版: 2015-10-08  

嗜铁钩端螺旋菌(Leptospirillum ferriphilum)gyrB基因的PCR扩增、克隆与序列分析

PCR Amplification, Cloning and Sequence Analysis of gyrB Gene with a Pair of Degenerate Primer from Leptospirillum ferriphilum
作者单位
1 湖南科技大学生命科学学院, 湖南 湘潭410201
2 中南大学资源加工与生物工程学院, 湖南 长沙410083
摘要
根据GenBank 和ICB 数据库中gyrB蛋白氨基酸序列的两个保守区域TPGMYIG和QRY(F)KGLGEM设计简并引物, 以L. ferriphilum菌株YSK基因组DNA为模板, PCR扩增出获得大小约为2.2 kb的DNA片段。序列分析表明, 菌株YSK的扩增片断的开放阅读框(ORF)能够推导出一个编码分子量约为82.24 kD、氨基酸数目为731个的蛋白质片断。这个氨基酸序列与所研究的gyrB蛋白氨基酸序列显示出高度的同源性, 尤其是与菌株AMC_Cont91的gyrB蛋白氨基酸序列的相似性高达92 %, 而与M. xanthus的gyrB的相似性最低, 仅为37 %。基于氨基酸序列的同源性及其所预测的蛋白质大小, 可以推断出该扩增片段属于gyrB基因。
Abstract
According to two conserved regions PGMYIG and QRY(F)KGLGEM of the amino acid sequences of the DNA gyrase subunit B proteins of bacteria from Genbank and ICB database, a pair of degenerate primer was designed to amplify the gyrB gene from L. ferriphilum strain YSK. Sequence analysis indicated that the ORF of PCR products from L. ferriphilum strain YSK could potentially encode a protein fragment of 731 amino acids with a molecular mass of 82.24 kDa. The deduced amino acid sequence of the ORF displayed strong homology to all gyrB proteins used in this study. The highest score was found with the product of AMC_Cont91 (92 % similarity). The similarity to the smallest G. sulfurreducens protein gyrB was 37 %. On the basis of the amino acid sequence homology and the predicted protein size, we concluded that the strain YSK ORF was the gyrB gene.

高健, 康健, 邱冠周. 嗜铁钩端螺旋菌(Leptospirillum ferriphilum)gyrB基因的PCR扩增、克隆与序列分析[J]. 激光生物学报, 2011, 20(3): 367. GAO Jian, KANG Jian, QIU Guan-zhou. PCR Amplification, Cloning and Sequence Analysis of gyrB Gene with a Pair of Degenerate Primer from Leptospirillum ferriphilum[J]. Acta Laser Biology Sinica, 2011, 20(3): 367.

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