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纸质出版:2008
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郝坡,刘北忠,欧阳峰,王东生,刘畅,钟梁,金丹婷,王翀.糖皮质激素受体结合域诱饵表达载体的构建及鉴定[J].陆军军医大学学报(原第三军医大学学报),2008,30(03):216-219.
HAO Po,LIU Bei-zhong,OUYANG Feng,WANG Dong-sheng,LIU Chang,ZHONG Liang,JIN Dan-ting,WANG Chong.Construction and identification of bait expression vector of glucocorticoid receptor binding domain[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(03):216-219.
郝坡,刘北忠,欧阳峰,王东生,刘畅,钟梁,金丹婷,王翀.糖皮质激素受体结合域诱饵表达载体的构建及鉴定[J].陆军军医大学学报(原第三军医大学学报),2008,30(03):216-219. DOI:
HAO Po,LIU Bei-zhong,OUYANG Feng,WANG Dong-sheng,LIU Chang,ZHONG Liang,JIN Dan-ting,WANG Chong.Construction and identification of bait expression vector of glucocorticoid receptor binding domain[J].J Amry Med Univ (J Third Mil Med Univ),2008,30(03):216-219. DOI:
目的 构建糖皮质激素受体(glucocorticoid receptor, GR)结合域的诱饵表达载体。 方法 RT-PCR扩增GR结合域,克隆入pMD18-T,测序正确后,再亚克隆入诱饵载体pGBKT7中,再把构建好的诱饵载体pGBKT7-GR转化到酵母AH109细胞中,并用Western blot分析诱饵蛋白的表达情况,同时检测诱饵蛋白的毒性和自激活作用。 结果 成功扩增了GR结合域,并分别成功克隆到pMD18-T和pGBKT7中,测序结果正确。诱饵载体成功转化到酵母AH109细胞中,无毒性和自激活作用,Western blot分析结果也证实了酵母细胞高表达诱饵蛋白。 结论 成功构建了GR结合域的酵母诱饵表达载体。
Objective To construct the bait expression plasmid pGBKT7-GR of glucocorticoid receptor (GR) binding domain. Methods The fragments of GR binding domain was amplified by RT-PCR
and then was cloned into pMD18-T. After being verified by sequencing
it was subcloned into the bait expression vector pGBKT7. Then the bait vector pGBKT7-GR was transformed into AH109 yeast cells and the expression of the bait protein was analyzed by Western blot. Toxicity and self-activation of the bait protein were detected. Results GR binding domain was amplified and cloned into pMD18-T and pGBKT7 successfully. The bait vector was transformed into AH109 yeast cells successfully
without toxicity or self-activation. The expression of the bait protein was confirmed by Western blot. Conclusion The successful construction of bait expression vector of glucocorticoid receptor binding domain lays the foundation for constructing small molecule ligand yeast three-hybrid system.
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