华北农学报 ›› 2020, Vol. 35 ›› Issue (5): 214-219. doi: 10.7668/hbnxb.20191066

• 资源环境·植物保护 • 上一篇    下一篇

我国主要流行的4种血清型禽腺病毒-Ⅰ群Fiber蛋白的交叉中和研究

栾庆东1, 孙举2, 刘均华3, 张本清4, 尹燕博1, 王建琳1   

  1. 1. 青岛农业大学 动物医学院, 山东 青岛 266109;
    2. 青岛博隆基因工程有限公司, 山东 青岛 266021;
    3. 青岛市农业行政执法支队, 山东 青岛 266071;
    4. 青岛市黄岛区农业农村局, 山东 青岛 266400
  • 收稿日期:2020-05-14 出版日期:2020-10-28
  • 通讯作者: 尹燕博(1963-),男,山东泰安人,教授,博士,主要从事动物病原致病机制及流行病学研究;王建琳(1976-),女,山西长治人,副教授,博士,主要从事禽类传染病致病机制及防控研究。
  • 作者简介:栾庆东(1994-),男,山东济南人,在读硕士,主要从事家禽传染病诊断方法的研究。
  • 基金资助:
    国家重点研发计划项目(2016YFD0500800)

Cross Neutralization of Fiber Protein Against Four Kinds of Serotype fowl adenovirus Prevalent in China

LUAN Qingdong1, SUN Ju2, LIU Junhua3, ZHANG Benqing4, YIN Yanbo1, WANG Jianlin1   

  1. 1. College of Veterinary Medicine, Qingdao Agricultural University, Qingdao 266109, China;
    2. Bolong Gene Therapeutic Biological Products Project, Qingdao 266021, China;
    3. Agricultural Administrative Law Enforcement Branch, Qingdao 266071, China;
    4. Agticultural and Rural Bureau of Huangdao District, Qingdao 266400, China
  • Received:2020-05-14 Published:2020-10-28

摘要: 为确定FAdV-4、FAdV-8a、FAdV-8b、FAdV-11这4个血清型FAdV间存在的交叉保护是否与病毒的纤维蛋白(Fiber)有关,制备4个血清型FAdV Fiber蛋白抗血清,在细胞水平研究其对本血清型和其他3种血清型病毒感染的中和作用。针对4个血清型FAdV的Fiber基因设计引物,进行PCR扩增,构建pCold-FAdV-Fiber重组质粒,分别转入BL21(DE3)感受态细胞,经IPTG诱导表达,通过SDS-PAGE和Western Blot鉴定表达产物;表达产物纯化后免疫SPF鸡,采集抗血清,通过LMH细胞研究不同血清型Fiber蛋白血清对各血清型FAdV感染的中和作用。PCR扩增结果显示,FAdV-4、FAdV-8a、FAdV-8b、FAdV-11 Fibe r基因产物大小分别为1 440,1 575,1 566,1 719 bp;重组菌在16℃培养条件下,1.0 mmol/L的IPTG诱导24 h可溶性重组蛋白的表达量最高;SDS-PAGE和Western Blot结果显示,FAdV-4、FAdV-8a、FAdV-8b、FAdV-11表达的蛋白分子量大小分别为102,107,107,112 ku;中和结果显示,FAdV-4、FAdV-8b和FAdV-11 Fiber蛋白抗血清能中和自身血清型FAdV对细胞的感染,不能中和其他血清型FAdV的感染,而FAdV-8a Fiber蛋白抗血清不能中和这4种血清型中的任何一种血清型FAdV的感染。综上,Fiber蛋白与4个血清型FAdV全病毒之间存在的交叉保护无直接关系。

关键词: FAdV, Fiber蛋白, 原核表达, 交叉中和

Abstract: To certain the effect of Fiber on the cross-neutralization among FAdV-4,FAdV-8a,FAdV-8b,FAdV-11,antisera against Fiber proteins of four Serotype fowl adenovirus were prepared to study the protective effect on the infection of fowl adenovirus with different serotypes at the LMH cell in the study. The recombinant plasmids of pCold-FAdV-Fiber were constructed after fiber gene was amplified by PCR,and the plasmids were transfected into BL21(DE3) and Fiber proteins were expressed. The products expressed were purified after identified by SDS-PAGE and Western Blot. SPF chickens were immunized with purified Fiber proteins and antisera were collected. The neutraliztion of antisera against Fiber proteins and different fowl adenovirus infection were studied on the LMH cells.The results showed that the size of FAdV-4,FAdV-8a,FAdV-8b,FAdV-11 PCR fragment were 1 440,1 575,1 566,1 719 bp differently,and the molecular weight of the proteins expressed were 102,107,107,112 ku. Antisera against Fibers of FAdV-4,FAdV-8b and FAdV-11 only neutralized infection of the virus with same serotype,and antiserum against Fibers of serotype FAdV-8a could not protect the LMH cells from infection of the virus with these four serotype. Comprehensively,Fiber proteins were not directly related to the cross-protection between the four serotypes FAdV viruses.

Key words: FAdV, Fiber protein, Prokaryotic expression, Cross neutralization

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引用本文

栾庆东, 孙举, 刘均华, 张本清, 尹燕博, 王建琳. 我国主要流行的4种血清型禽腺病毒-Ⅰ群Fiber蛋白的交叉中和研究[J]. 华北农学报, 2020, 35(5): 214-219. doi: 10.7668/hbnxb.20191066.

LUAN Qingdong, SUN Ju, LIU Junhua, ZHANG Benqing, YIN Yanbo, WANG Jianlin. Cross Neutralization of Fiber Protein Against Four Kinds of Serotype fowl adenovirus Prevalent in China[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(5): 214-219. doi: 10.7668/hbnxb.20191066.

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