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Published on: July 16, 2013
[Interaction of connexin 26 with the C-terminal of neuroendocrine specific protein]
Zi-An Xiao1, Ding-Hua Xie, Liang-Qun Huang
1Department of Otolaryngology and Hearing Research Laboratory, Second Xiangya Hospital, Central South University, Changsha, China. xiaozian@hotmail.com
Objective:
To screen and identify the interactive proteins with connexin 26 (Cx26) by the yeast two hybrid technique.
Methods:
The whole coding region of Cx26 (GJB2) gene was amplified from normal human genomic DNA by polymerase chain reaction (PCR). The "bait" Cx26 was then subcloned into the vector pGBKT7 plasmid of the MatchMaker Gal4 Two-Hybrid System 3 as a target to screen its interactive proteins ("prey") from the human fetal brain eDNA library by the yeast two hybrid technique. The false positive clones were discarded from the preys by one to one yeast two hybrid method between Cx26 and the preys. The DNAs of the preys were sequenced and BLAST analyzed against the GenBank, and also underwent other bioinformatics analysis.
Results:
The insert of one positive clone contained 145 amino acids residues that was identical to the C-terminal of the neuroendocrine specific protein (NSP) and the open reading frame of the insert was correct.
Conclusion:
Cx26 is interacted with the C-terminal of NSP. NSP may participate in the process of Cx26 transportation, assembling the connexon, or influencing the functions of its connexons.
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