中文摘要：

目的构建人巨细胞病毒UL49基因的诱饵表达载体，以利于筛选与pUL49相互作用的蛋白。方法PCR扩增UL49全序列，克隆入pGBKT7，将构建好的pGBKT7-UL49转化到酵母细胞AH109；用蛋白印迹法分析诱饵蛋白的表达，检测诱饵蛋白有无毒性和自激活效应。结果克隆成功pGBKT7-UL49；pG—BKT7-UL49成功转化到AH109，转化细胞AH109[pGBKT7-UL49]表达诱饵蛋白pUL49，pUL49对转化细胞无细胞毒性、无自激活。结论成功构建了酵母诱饵表达载体pGBKT7-UL49，为进-步筛选与pUL49相互作用的蛋白提供了实验基础。

英文摘要：

[ Objective ] To construct the bait expression vector pGBKT7-UL49 of human cytomegalovirus UL49 gene coding protein for screening the target proteins interacting with the bait protein pUL49 through the yeast twohybrid system. [Methods] The fragments of UIA9 was amplified by PCR, and then cloned into the bait expression vector pGBKT7. The bait vector pGBKTT-UL49, being verified by sequencing, was transformed into AH109 yeast cells. Then the bait protein pUL49 was analyzed by Western Blotting. Toxicity and self-activation of the bait protein were detected by cultured in different SD. [ Results ] UL49 was amplified and cloned into pGBKT7 successfully. The vector pGBKT7-UL49 was transformed into AH109 as well and those cells exhibited neither toxicity nor self-activation. The expression of the bait protein pUL49 was detected by Western Blotting. [ Conclusion ] The bait expression vector of UL49 was constructed successfully, which layed the foundation for screening target proteins interacting with the bait protein pUL49 using the yeast two-hybrid technique.