[Interaction of connexin 26 with the C-terminal of neuroendocrine specific protein].
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.


CONCLUSIONS
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.
Xiao
Zi-An
ZA
Department of Otolaryngology and Hearing Research Laboratory, Second Xiangya Hospital, Central South University, Changsha, China. xiaozian@hotmail.com
Xie
Ding-Hua
DH
Huang
Liang-Qun
LQ
Shi
Xiao-Liu
XL
Xia
Kun
K
Yang
Shu
S
Xia
Jia-Hui
JH
D000429:Alcohol Oxidoreductases; D000595:Amino Acid Sequence; D001483:Base Sequence; D001921:Brain / Q000378:metabolism*; D000072259:Connexin 26; D017630:Connexins / Q000737:chemistry* / Q000235:genetics; D004268:DNA-Binding Proteins / Q000737:chemistry; D005333:Fetus; D015723:Gene Library; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics; D010750:Phosphoproteins / Q000737:chemistry; D011485:Protein Binding; D025941:Protein Interaction Mapping; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000235:genetics
D017630:Connexins; D004268:DNA-Binding Proteins; C000607042:GJB2 protein, human; D009419:Nerve Tissue Proteins; D010750:Phosphoproteins; C081781:RTN1 protein, human; D000072259:Connexin 26; D000429:Alcohol Oxidoreductases; C096794:C-terminal binding protein