O139群霍乱弧菌毒力岛区域分子特征分析  

Molecular characters of Vibrio cholerae pathogenicity island of Vibrio cholerae O139

作　　者：王颖芳[1] 段广才[2,3] 谢婧[2] 郗园林[2] 范清堂[3] 

机构地区：[1]河南科技大学预防医学教研室,洛阳471003 [2]郑州大学公共卫生学院 [3]河南省分子医学重点学科开放实验室

出　　处：《中国公共卫生》2009年第1期58-60,共3页Chinese Journal of Public Health

摘　　要：目的比较O139群霍乱弧菌国际标准株MO45和O1群E l Tor型霍乱弧菌国际标准株N16961基因组中毒力岛区域分子特征。方法从MO45基因组中筛选含有毒力岛上游的VC0815基因及其上游基因片段并克隆到pNEB193中命名为pNEBVC081545,绘制pNEBVC081545的限制性酶切图谱并与N16961同段序列限制性酶切图谱进行比较。在pNEBVC081545中选择VC0815基因上游的一段侧序列进行亚克隆,测序后与N16961同段序列比较。结果pNEBVC081545限制性酶切图谱中VC0815基因上游序列图谱中第1个碱基位置是限制性内切酶位点为KpnⅠ,第732位为SalⅠ,第819位为HpaⅠ,第1578位为BglⅡ,第1600位为H indⅢ,第1943位为XbaⅠ,第3247位为EcoRⅠ,此段序列限制性内切酶图谱与N16961同段序列限制性酶切图谱完全相同。对SalⅠ和EcoRⅠ之间的序列进行亚克隆,并对该段序列测序,测序结果表明该段核酸序列与N16961中同段序列完全相同。结论O139霍乱弧菌可能是E l Tor霍乱弧菌O抗原突变而产生的菌群。Objective To compare molecular character of Vibrio cholerae pathogenicity island （VPI） of Vibrio cholerae O139 international reference strain MO45 and E1 Tot biotype of Vibrio cholerae international reference strain N16961. Methods Segments including VC0815 gene in the upriver of VPI and it＇s upriver segment from MO45 were screened and cloned to pNEBVC081545. Restriction map of pNEBVC081545 was drawn and compared with the map of the same segment of N16961. The pNEBVC081545 was subcloned and analyzed. Results There were several restriction enzyme sites in the upriver segment of VC0815 gene in the restriction map of pNEB -VC081545 including Kpn Ⅰ at the 1st base pair,Sal Ⅰ at the 732nd base pair,Hpa Ⅰ at the 819th base pair, Bgl Ⅱ at the 1578th base pair, HindⅢ at the 1600th base pair,Xba Ⅰ at the 1943rd base palr,EcoR Ⅰ at the 3274th base pair. The restriction map of the segment was the same to the segment of N16961. The segment between Sal Ⅰ and EcoR Ⅰ subcloned was the same to N16961. Conclusion The result of the study supports the view that Vibrio cholerae O139 maybe results from the mutation of the O antagen of E1 Tor biotype of Vibrio cholerae.

关 键 词：霍乱弧菌 毒力岛 限制性酶切图谱 测序 

分 类 号：R183[医药卫生—流行病学]