Construction of GJB2 mutations common in Chinese EGFP fusion protein vectors / 临床耳鼻咽喉头颈外科杂志
Lin chuang er bi yan hou ke za zhi
; (24): 724-727, 2009.
Article
in Zh
| WPRIM
| ID: wpr-434256
Responsible library:
WPRO
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.
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WPRIM
Language:
Zh
Journal:
Lin chuang er bi yan hou ke za zhi
Year:
2009
Type:
Article