酵母双杂交诱饵表达载体pGBKT7-Tiam1/C1199的构建  

Construction of yeast two-hybrid bait expression vector pGBKT7-Tiam1/C1199

作　　者：黄静 陈龙华[2] 丁轶[2] 

机构地区：[1]广东医科大学附属医院肿瘤中心,广东湛江524001 [2]南方医科大学南方医院放疗科,广东广州510515

出　　处：《广东医学院学报》2016年第4期357-361,共5页Journal of Guangdong Medical College

基　　金：国家自然科学基金(No.81502532);广东省自然科学基金(No.2014A030310101);湛江市科技计划项目(No.2014A06005);广东医学院科研基金(No.Z2014004)

摘　　要：目的 构建人T细胞淋巴瘤侵袭转移诱导因子1 （T lymphoma invasion and metastasis 1, Tiam1）诱饵表达载体。方法 通过PCR扩增获得人Tiam/C1199基因片段，克隆至pGBKT7酵母诱饵表达载体，经测序鉴定后，转化酵母菌株AH109，分析Tiam1蛋白在酵母菌株中表达情况。结果 成功扩增了Tiam1/C1199基因片段，并克隆入pGBKT7中。诱饵表达载体成功转化到酵母菌株AH109中，且Tiam1诱饵蛋白表达良好，无毒性及自激活活性。结论 成功构建了pGBKT7-Tiam1/C1199酵母诱饵蛋白表达载体。Objective To construct the bait expression plasmid pGBKT7-T lymphoma invasion and metastasis 1 （Tiam 1）/C 1199. Methods The Tiam 1/C 1199 fragment was amplified by PCR and cloned into the bait vector pGBKT7. After identified by DNA sequencing, the recombinant plasmid was transformed into AH109 yeast cells. Tiaml expression was then analyzed. Results The Tiaml/C1199 fragment was successfully amplified and cloned into pGBKT7. The bait expression vector was transformed into AH109 yeast cells. Tiaml protein was expressed in the bait vector, without toxicity and self- activation. Conclusion The bait plasmid pGBKT7-Tiaml/Cl199 is successfully constructed using yeast two-hybrid technique.

关 键 词：TIAM1 诱饵蛋白 酵母双杂交 

分 类 号：R587.1[医药卫生—内分泌]