Construction and identification of bait recombinant vector of pGBKT7-tumor necrosis factor-alpha converting enzyme cytoplasmic tail in the yeast two-hybrid system.
- Author:
Xin LIU
1
;
Feng-dan WANG
;
Jian-xing WANG
;
Xue-fei ZHAO
;
Jing YU
;
Xiao-guang YU
Author Information
- Publication Type:Journal Article
- MeSH: ADAM Proteins; genetics; metabolism; ADAM17 Protein; Animals; Cloning, Molecular; DNA, Complementary; Male; Mice; Mice, Inbred BALB C; Plasmids; RNA; isolation & purification; Reverse Transcriptase Polymerase Chain Reaction; Testis; metabolism; Two-Hybrid System Techniques; Yeasts; metabolism
- From: National Journal of Andrology 2007;13(3):229-232
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct the bait vector pGBKT7-TACEc (cytoplasmic tail of tumor necrosis factor-alpha converting enzyme) of Macthmaker GAL4 Two-hybrid System 3, and to test whether it has self-activation and toxic action.
METHODSTACEc gene was amplified by RT-PCR from the mouse testis, and the EcoRI and BamHI sites were introduced into it. The TACEc gene, after sequenced, was cloned into pGBKT7. Self-activation and toxic action of the recombination vector pGBKT7-TACEc was tested.
RESULTSThe pGBKT7-TACEc vector was successfully constructed and proved of no self-activation and toxic action.
CONCLUSIONThe pGBKT7-TACEc can be applied to the screening of the mouse testis cDNA library in the yeast two-hybrid system.
