Ⅱ群禽腺病毒五邻体蛋白单因子血清的制备

›› 2013, Vol. 40 ›› Issue (10): 16-20.

Ⅱ群禽腺病毒五邻体蛋白单因子血清的制备

刘雪美, 秦建如, 李育豪, 曹伟胜

华南农业大学兽医学院, 农业部兽用疫苗创制重点实验室, 广东广州 510642

收稿日期:2013-02-18 出版日期:2013-10-20 发布日期:2013-12-19
通讯作者: 曹伟胜 E-mail:weishengcao@yahoo.com
作者简介:刘雪美(1986-),女,山东人,硕士生,研究方向:动物传染病。秦建如(1989-),女,河南人,硕士生,研究方向:动物传染病。刘雪美和秦建如对本文具有同等贡献,并列为第一作者。
基金资助:
农业部肉鸡产业技术体系(nycytx-42-G3-03);广东省科技计划项目(2012A020100001)。

Preparation of Uni-factor Serum against Penton Protein of Group Ⅱ Avian Adenovirus

LIU Xue-mei, QIN Jian-ru, LI Yu-hao, CAO Wei-sheng

Key Laboratory for Animal Vaccine Development, Ministry of Agriculture, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642, China

Received:2013-02-18 Online:2013-10-20 Published:2013-12-19

摘要/Abstract

摘要： 通过PCR方法扩增得到Ⅱ群禽腺病毒火鸡出血性肠炎病毒(HEV)的五邻体(penton)基因,并构建了原核表达载体pET-32a-penton,将其转化至E.coil BL21中,通过诱导表达,以切胶回收的方式纯化蛋白,获得约70 ku的重组融合蛋白,Western blotting检测纯化后的重组蛋白具有较好的抗原性。将纯化后的重组蛋白免疫新西兰大白兔,制备了五邻体蛋白单因子血清,间接ELISA检测结果显示所制备的抗血清效价大于1:40000。本试验获得的高效单因子血清,为Ⅱ群禽腺病毒特异性检测方法的研究奠定了基础。

关键词: Ⅱ群禽腺病毒; 五邻体蛋白; 单因子血清

Abstract: A pair of primers was designed based on the genome sequence of hemorrhagic enteritis virus VAS strain, the complete gene of penton with a size of 1347 bp was amplified by PCR, and then cloned into plasmid expression vector pET-32a (+), the obtained positive clone was named pET-32a-penton. The pET-32a-penton was transformed into E.coli BL21, after inducing expression and purification of the protein in gel slices, about 70 ku recombinant fusion protein was obtained and used to immunize New Zealand White rabbits to prepare uni-factor serum against penton protein. Indirect ELISA was used to detect the antibody titer. The results showed that the antisera titer was greater than 1:40000. The method to get the efficient uni-factor serum was simple and low-cost, which laid a foudation for detection of group Ⅱ avian adenovirus.

Key words: group Ⅱ avian adenovirus; penton protein; uni-factor serum

中图分类号:

S852.5

刘雪美, 秦建如, 李育豪, 曹伟胜. Ⅱ群禽腺病毒五邻体蛋白单因子血清的制备[J]. , 2013, 40(10): 16-20.

LIU Xue-mei, QIN Jian-ru, LI Yu-hao, CAO Wei-sheng. Preparation of Uni-factor Serum against Penton Protein of Group Ⅱ Avian Adenovirus[J]. , 2013, 40(10): 16-20.

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