黄瓜花叶病毒运动蛋白基因及其缺失突变体在大肠杆菌中的表达和表达产物的纯化

EXPRESSION AND PURIFICATION OF MOVEMENT PROTEIN AND ITS TWO MUTANTS OF CUCUMBER MOSAIC VIRUS

利用ＣＭＶＦｎｙ 株系ＲＮＡ３ 全长ｃＤＮＡ 克隆， 构建了运动蛋白（ＭＰ） 基因５′端缺失突变体和３′端缺失突变体的原核表达载体。ＳＤＳ－ ＰＡＧＥ 分析表明， 经ＩＰＴＧ 诱导， ＭＰ 基因及其２ 种缺失突变体均能在大肠杆菌ＢＬ２１（ＤＥ３） ｐＬｙｓＳ中高效表达。利用分离包含体的方法， 提纯了全长的及Ｃ 端缺失的ＭＰ。光密度扫描分析表明， 提纯产物的纯度达９６ ．６ ％ 。

Two expression vectors containing 5′ regional deletion and 3′ regional deletion of cucumber mosaic virus movement protein (MP) gene were constructed respectively. With IPTG induction, the full length of MP and its two mutants were overexpressed in Escherichia coli BL21(DE3) pLysS respectively. Using a method to separate the inclusion bodies from the rest of the bacterial proteins by centrifugation, the MP and its C terminal deletion mutant were purified. The purity was about 96.6% estimated by Shimadzu 910 scanning.