转座子IS10新插入位点的序列分析

Sequence analysis of a novel insertion site of transposon IS10.

作者信息

Xiang Tai-He, Wang Li-Lin, Wang Hui-Zhong

机构信息

School of Life Sciences, Hangzhou Normal College, Hangzhou 310036, China.

出版信息

Yi Chuan Xue Bao. 2006 Nov;33(11):1047-52. doi: 10.1016/S0379-4172(06)60141-8.

DOI:10.1016/S0379-4172(06)60141-8

PMID:17112977

Abstract

A sacB mutant was obtained by transposon IS10 inactivation of a plasmid pXT3sacB carrying the sacB gene. Sequencing of this mutant plasmid DNA (GenBank accession No. AY580883.1) showed that the IS10 flanking the 22 bp inverted repeats were 5'-CTGAGAGATCCCCTCATAATTT-3' and 5'-AAATCATTAGGGGATTCATCAG-3', which were the similar to those published in reports previously. However, the target sequence adjacent to IS10 was 5'-TGCTTGGTT-3' instead of the previously reported 5'-NGCTNAGCN-3'. To our knowledge, this is the first report on the novel insertion site of IS10. In addition, Southern blot hybridization confirmed that the mobile IS10 originated from the chromosomal DNA of the host strain Escherichia coli DH5alpha and that there were two copies in the DH5alpha genome.

摘要

通过转座子IS10使携带sacB基因的质粒pXT3sacB失活，获得了一个sacB突变体。对该突变体质粒DNA进行测序（GenBank登录号为AY580883.1），结果显示，位于22 bp反向重复序列两侧的IS10分别为5'-CTGAGAGATCCCCTCATAATTT-3'和5'-AAATCATTAGGGGATTCATCAG-3'，这与之前报道中的序列相似。然而，与IS10相邻的靶序列是5'-TGCTTGGTT-3'，而不是之前报道的5'-NGCTNAGCN-3'。据我们所知，这是关于IS10新插入位点的首次报道。此外，Southern杂交证实，可移动的IS10来源于宿主菌株大肠杆菌DH5α的染色体DNA，并且在DH5α基因组中有两个拷贝。