Construction and identification of bait recombinant vector of pGBKT7-tumor necrosis factor-alpha converting enzyme cytoplasmic tail in the yeast two-hybrid system / 中华男科学杂志

ABSTRACT

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>The pGBKT7-TACEc vector was successfully constructed and proved of no self-activation and toxic action.</p><p><b>CONCLUSION</b>The pGBKT7-TACEc can be applied to the screening of the mouse testis cDNA library in the yeast two-hybrid system.</p>