畜牧与兽医2018，Vol.50Issue(2)：83-85,3.
禽安卡拉病毒Hexon蛋白多克隆抗体的制备
Preparation of polyclonal antibody against avian Ankara virus Hexon protein
刘兰 1卢渊录 1梁雄燕 1蔡宇威 1顾玉芳1
作者信息
- 1. 长江大学动物科学学院,湖北荆州434025
- 折叠
摘要
Abstract
Based on the group Ⅰ fowl adenovirus serotype 4 (FADV-4) genome sequence,a pair of specific primers were designed to amplify the hexon gene by PCR,and then cloned into the pET-28a vector named pET-28a-Hexon.The recombinant plasmid was transformed into competent cells BL21 induced with IPTG for expression.The results showed that 49 ku of the fusion proteins was obtained following an SDS-PAGE analysis.Expressions induced with different IPTG concentrations,temperatures and durations suggested the best IPTG induction concentration to be 1 mmol/L,the optimum temperature to be 37℃,and the optimal duration of time to be 4 h.Western-bolt analysis showed that the recombinant fusion proteins reacted specifically with the FAdV-4 positive serum.The purified recombinant proteins were inoculated in BALB/c mice to prepare polyclonal antibody against hexon protein.The titer of the polyclonal antibody was found to be higher than 1 ∶ 64 000,assayed by indirect ELISA.The high efficiency single factor serum prepared in this experiment laid the foundation for the study on the specific detection of serotype 4 adenovirus.关键词
Ⅰ群4型禽腺病毒/六邻体蛋白/多克隆抗体Key words
group Ⅰ fowl adenovirus serotype 4/hexon protein/polyclonal antibody分类
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刘兰,卢渊录,梁雄燕,蔡宇威,顾玉芳..禽安卡拉病毒Hexon蛋白多克隆抗体的制备[J].畜牧与兽医,2018,50(2):83-85,3.
