Construction of GJB2 mutations common in Chinese EGFP fusion protein vectors.
- Author:
Yanping ZHANG
1
;
Yuanding ZHANG
;
Lina LI
;
Lei MA
;
Yurui SUN
;
Zonglin ZHANG
;
Jinwei LIU
;
Huiyan DENG
;
Wei ZHU
Author Information
1. Department of Otolaryngology, the Second Affiliated Hospital of General Hospital of Chinese PLA, Beijing, 100091, China. yzhan28@163.com
- Publication Type:Journal Article
- MeSH:
Asian Continental Ancestry Group;
genetics;
Connexin 26;
Connexins;
genetics;
Genetic Vectors;
Green Fluorescent Proteins;
genetics;
Humans;
Sequence Deletion
- From:
Journal of Clinical Otorhinolaryngology Head and Neck Surgery
2009;23(16):724-727
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To construct GJB2 gene mutations common in Chinese EGFP fusion protein vectors, and to search for better way to study the mechanism of deletion mutations in GJB2 gene.
METHOD:Non-fusion protein vectors of 235delC, 299-300 del AT and 176 del 16 bp were first made by point mutation 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 mutations were inserted into pEGFP-N1 vector. Sequencing was used to verify the validity of the fusion protein vectors. HEK293 cells were transfected with the recombinant DNA samples by the liposome complex method.
RESULT:The recombined plasmids were highly expressed in HEK293 cells. Green fluorescence signals 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.
