Related Experiment Video
Updated: Jun 17, 2026
Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
Published on: March 28, 2018
[Construction of GJB2 mutations common in Chinese EGFP fusion protein vectors]
Yanping Zhang1, Yuanding Zhang, Lina Li
1Department of Otolaryngology, the Second Affiliated Hospital of General Hospital of Chinese PLA, Beijing, 100091, China. yzhan28@163.com
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.
