[Construction and identification of bait recombinant vector of pGBKT7-tumor necrosis factor-alpha converting enzyme cytoplasmic tail in the yeast two-hybrid system].
OBJECTIVE
To 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.


METHODS
TACEc 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.


RESULTS
The pGBKT7-TACEc vector was successfully constructed and proved of no self-activation and toxic action.


CONCLUSIONS
The pGBKT7-TACEc can be applied to the screening of the mouse testis cDNA library in the yeast two-hybrid system.
Liu
Xin
X
Department of Biochemistry and Molecular Biology, Harbin Medical University, Harbin, Heilongjiang 150086, China.
Wang
Feng-dan
FD
Wang
Jian-xing
JX
Zhao
Xue-fei
XF
Yu
Jing
J
Yu
Xiao-guang
XG
D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D000072198:ADAM17 Protein; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D012313:RNA / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013737:Testis / Q000378:metabolism*; D020798:Two-Hybrid System Techniques; D015003:Yeasts / Q000378:metabolism
D018076:DNA, Complementary; D012313:RNA; D051722:ADAM Proteins; D000072198:ADAM17 Protein