汉滩病毒糖蛋白G2酵母双杂交诱饵载体的构建及初步鉴定

目的利用Ras募集系统(RRS),构建并鉴定含汉滩病毒囊膜糖蛋白G2的诱饵载体。方法将汉滩病毒76-118株囊膜糖蛋白G2基因与Ras基因连接,构建嵌合基因Ras-G2及Ras-G2’(G2’无前导肽序列)。将两个嵌合基因分别克隆入酵母表达载体pMet25,并转化至酵母温度敏感株cdc25-2,检测其对RRS系统的激活作用。结果限制性内切酶酶切鉴定表明,Met-G2和Met-G2’载体构建正确。激活试验结果为阴性。结论Met-G2和Met-G2’载体可用于从cDNA文库中筛选汉滩病毒受体。

Construction and identification of the bait recombinant vector of Hantaan virus (HTNV) glycoprotein G2

To construct and primarily identify the bait recombinant vector of Hantaan virus (HTNV) by using the Ras recruitment system (RRS), the glycoproteins of HTNV-G2 and HTNV-G2'(mutant of G2 without signal sequence) were ligated to Ras gene at the C-terminal of Ras, and then these two chimeric genes were cloned into yeast expression vector pMet25 of RRS. The recons were transfected into the temperature-sensitive strain cdc25-2 and tested for their effect on the activation of the recons to RRS. It was found the bait recombinant vectors Met-G2 and Met-G2'were constructed successfully by restriction enzyme digestion analysis, but the activation testing showed a negative result. It concludes that the Met-G2 and Mey-G2' vectors can be used to screen the receptors of HTNV from cDNA library.