title: [Construction of GJB2 mutations common in Chinese EGFP fusion protein vectors].
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.
  
  
  METHODS
  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.
  
  
  RESULTS
  The recombined plasmids were highly expressed in HEK293 cells. Green fluorescence signals were distributed uniformly in cytoplasm.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Otolaryngology, the Second Affiliated Hospital of General Hospital of Chinese PLA, Beijing, 100091, China. yzhan28@163.com
authors.forename: Yanping
authors.initials: Y
authors.lastname: Zhang
authors.forename: Yuanding
authors.initials: Y
authors.lastname: Zhang
authors.forename: Lina
authors.initials: L
authors.lastname: Li
authors.forename: Lei
authors.initials: L
authors.lastname: Ma
authors.forename: Yurui
authors.initials: Y
authors.lastname: Sun
authors.forename: Zonglin
authors.initials: Z
authors.lastname: Zhang
authors.forename: Jinwei
authors.initials: J
authors.lastname: Liu
authors.forename: Huiyan
authors.initials: H
authors.lastname: Deng
authors.forename: Wei
authors.initials: W
authors.lastname: Zhu
mesh_terms: D044466:Asian People / Q000235:genetics; D000072259:Connexin 26; D017630:Connexins / Q000235:genetics*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006801:Humans; D017384:Sequence Deletion
chemical_list: D017630:Connexins; C000607042:GJB2 protein, human; C485184:enhanced green fluorescent protein; D000072259:Connexin 26; D049452:Green Fluorescent Proteins