Bioinformatics analysis and construction of eukaryotic expression plasmid of Cx50 V64G mutation
- VernacularTitle:Cx50 V64G突变的生物信息学分析及其真核表达载体的构建
- Author:
Ping, LIU
;
Ying, LIN
;
Yue-Ying, YANG
;
Jian-Qiu, ZHENG
;
Ying, HOU
;
Di, JIN
;
Xiao-Bo, FU
;
Hong-Mei, MA
- Publication Type:Journal Article
- Keywords:
Cx50;
bioinformatics analysis;
eukaryotic expression plasmid
- From:
International Eye Science
2007;7(5):1206-1208
- CountryChina
- Language:Chinese
-
Abstract:
AIM : To construct and analyze eukaryotic expression plasmid inserted by Cx50 with V64G mutation through bioinformatics software.METHODS: The full coding domain sequence of Cx50 with V64G mutation was acquired from the blood of patients with cataract and was cloned into pcDNA3.1 /Amp (+).The constructed plasmid was identified with PCR , enzyme digestion and sequencing. The analysis of Cx50 with V64G mutation was performed with bioinformatics software.RESULTS : Cx50 with V64G mutation was successfully amplified and its eukaryotic expression plasmid was constructed. Valine-64 is well conserved in the first extracellular loop of connexin 50 in different species and also in different human α -type gap junctional proteins.CONCLUSION : The successive reconstruction and verification of eukaryotic expression plasmid containing Cx50 with V64G mutation established the foundation for further studying the mechanism of cataract.
