Construction of GJB2 mutations common in Chinese EGFP fusion protein vectors
10.3969/j.issn.1001-1781.2009.16.002
- VernacularTitle:中国人常见GJB2基因突变表达载体的构建及鉴定
- Author:
Yanping ZHANG
;
Yuanding ZHANG
;
Lina LI
;
Lei MA
;
Yurui SUN
;
Zonglin ZHANG
;
Jinwei LIU
;
Huiyan DENG
;
Wei ZHU
- Publication Type:Journal Article
- Keywords:
connexin26;
mutation;
green fluorescent protein
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2009;(16):724-727
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct GJB2 gene mutaitons common in Chinese EGFP fusion protein vectors, and to search for better way to study the mechanism of deletion mutaitons in GJB2 gene. Method: Non-fusion protein vectors of 235delC, 299-300 del AT and 176 del 16 bp were first made by point mutaiton methods in vitro. Then expression part of the upper 3 mutations were amplified by PCR and the PCR products were cloned into TA cloning vector. After cutting by restriction enzymes EcoRI/BamHI, three deletion mutaions were inserted into pEG-FP-N1 vector. Sequencing was used to verify the validity of the fusion protein vectors. HEK293 cells were trans-fected with the recombinant DNA samples by the liposome complex method. Results The recombined plasmids were highly expressed in HEK293 cells. Green fluorescence singals were distributed uniformly in cytoplasm. Conclusion; GJB2 mutations common in Chinese EGFP fusion protein vectors were constructed successfully. It may provide a better way to explore the reasons of nonsyndromic hearing loss common in Chinese.
