小麦黄花叶病毒RNA2 28kDa蛋白基因的表达及表达产物抗血清的制备

Overexpression of 28?kDa Protein Encoded by Wheat Yellow Mosaic Virus RNA2 in \%E. coli\%

根据小麦黄花叶病毒（ Ｗ Ｙ Ｍ Ｖ） 核苷酸序列测定结果，将 Ｗ Ｙ Ｍ Ｖ Ｒ Ｎ Ａ２ 上的２８ ｋ Ｄａ 蛋白基因克隆到ｐ Ｅ Ｔ１１ａ 上，构建了原核表达载体ｐ Ｅ２８３９ 。 Ｓ Ｄ Ｓ Ｐ Ａ Ｇ Ｅ 分析表明，经 Ｉ Ｐ Ｔ Ｇ 诱导，２８ ｋ Ｄａ蛋白基因在大肠杆菌 Ｂ Ｌ２１（ Ｄ Ｅ３）ｐ Ｌｙｓ Ｓ 中得到高效表达。以含表达产物的凝胶为抗原，免疫家兔，首次制备了小麦黄花叶病毒 Ｒ Ｎ Ａ２ 蛋白特异性抗血清。

With polymerase chain reaction (PCR) a fragment encoding the 28 kDa protein of wheat yellow mosaic virus (WYMV) RNA2 was amplified and constructed into plasmid of pET11a for prokaryotic expression. BL21 (DE3) of \%E.coli\% transformed with the recombinant plasmid of pE2839 was induced to specifically express the 28 kDa protein in high level. The expressed 28 kDa protein was purified from SDS polyacrylamide gels and the antiserum against the protein was raised in rabbit. In Western blotting analysis, the antiserum reacted with the 28?kDa protein.