Overexpression of 28 kDa Protein Encoded by W heat Yellow M osaic Virus RNA2 in E．coli

SU Ning; HAN Cheng-Gui; LI Da-Wei; HOU Tie-Jun; GENG Yi-Meng; XU Jia-Lin; LIU Yi

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June 1999

SU Ning, HAN Cheng-Gui, LI Da-Wei, HOU Tie-Jun, GENG Yi-Meng, XU Jia-Lin and LIU Yi. Overexpression of 28 kDa Protein Encoded by W heat Yellow M osaic Virus RNA2 in E．coli[J]. Virologica Sinica, 1999, 14(3): 217-221.

Citation: SU Ning, HAN Cheng-Gui, LI Da-Wei, HOU Tie-Jun, GENG Yi-Meng, XU Jia-Lin, LIU Yi. Overexpression of 28 kDa Protein Encoded by W heat Yellow M osaic Virus RNA2 in E．coli .VIROLOGICA SINICA, 1999, 14(3) : 217-221.

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

1.
中国农业大学生物技术国家重点实验室，北京100094

摘要

根据小麦黄花叶病毒（ＷＹＭＶ）核苷酸序列测定结果，将ＷＹＭＶＲＮＡ２上的２８ｋＤａ蛋白基因克隆到ｐＥＴ１１ａ上，构建了原核表达载体ｐＥ２８３９。ＳＤＳＰＡＧＥ分析表明，经ＩＰＴＧ诱导，２８ｋＤａ蛋白基因在大肠杆菌ＢＬ２１（ＤＥ３）ｐＬｙｓＳ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ＲＮＡ２蛋白特异性抗血清。
- 小麦黄花叶病毒
- , 蛋白基因
- , 原核表达
- , 抗血清

Overexpression of 28 kDa Protein Encoded by W heat Yellow M osaic Virus RNA2 in E．coli

1.
State Key LaboraZory for Agrobiola-hnology，China Agricultural Oniz~rsizy，Beijing 100094

Abstract

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.
- Wheat yellow mosaic virus
- , Protein gene
- , Prokaryotic expression
- , Antiserum

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Overexpression of 28 kDa Protein Encoded by W heat Yellow M osaic Virus RNA2 in E．coli

1. State Key LaboraZory for Agrobiola-hnology，China Agricultural Oniz~rsizy，Beijing 100094

Abstract: 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.