安卡拉病毒贵州分离株Fiber2基因的克隆及序列分析  

Cloning and sequence analysis of Fiber2 gene of Ankara virus Guizhou strain

作　　者：方天 何玲 尹德晶 岳筠[2] 朱二鹏 文明[1,3] 程振涛 FANG Tian;HE Ling;YIN De-jing;YUE Jun;ZHU Er-peng;WEN Ming;CHENG Zhen-tao(College of Animal Science,Guizhou University,Guiyang 550025,China;Guizhou Provincial Center for Animal Disease Control and Prevention,Guiyang 550008,China;Guizhou Provincial Key Laboratory of Animal Disease and Veterinary Public Health,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025 [2]贵州省动物疫病预防控制中心,贵州贵阳550008 [3]贵州省动物疫病与兽医公共卫生重点实验室,贵州贵阳550025

出　　处：《江苏农业学报》2022年第6期1603-1611,共9页Jiangsu Journal of Agricultural Sciences

基　　金：贵州省科技计划项目[黔科合基础(2020)1Y409号];贵州省科技支撑计划项目[黔科合支撑(2021)一般161];贵州省优秀青年科技人才项目[黔科合平台人才(2021)5646]。

摘　　要：Fiber2蛋白是禽腺病毒血清4型(FAdV-4,又称安卡拉病毒)重要的免疫原蛋白,为深入了解FAdV-4贵州株Fiber2基因的遗传变异情况,本研究利用PCR技术扩增FAdV-4贵州晴隆株(GZ-QL)Fiber2基因,胶回收后将其连接至pMD19-T载体,转化入DH5α筛选阳性克隆质粒,经质粒PCR及双酶切鉴定后进行DNA测序,然后对测序结果进行生物信息学分析。结果显示,GZ-QL株Fiber2基因全长为1 440 bp,与四川分离的SCnj1601强毒株、北京分离的NIVD2强毒株Fiber2基因核苷酸同源性均为100.0%,与FAdV-4经典ON1株Fiber2基因核苷酸同源性为95.9%,两者位于相同进化分支。GZ-QL Fiber2蛋白共有33个氨基酸突变位点,其中,在第11~15位之间有5个氨基酸插入;蛋白质亲水性平均值为-0.031,其二级结构以无规则卷曲为主(占51.8%),无跨膜结构域或信号肽,抗原表位分布均匀,预测有8个优势抗原表位区域,且集中于N端。以上结果表明,FAdV-4贵州株GZ-QL Fiber2蛋白在禽腺病毒血清4型内相对保守且抗原性良好,具备成为优势抗原的潜力。Fiber2 protein is an important immunogenic protein of fowl adenovirus serotype 4(FAdV-4, also known as Ankara virus). In order to gain deep understanding of the genetic variation of Fiber2 gene of FAdV-4 Guizhou strain, PCR technology was used to amplify the Fiber2 gene of FAdV-4 Guizhou Qinglong strain(GZ-QL). After the gene was recycled from the separation gel, it was connected into pMD19-T vector and then converted into DH5α to screen positively cloned plasmids. After identification by plasmid PCR and dual-enzyme digestion, the DNA was sequenced, and then bioinformatic analysis of the sequencing results was conducted. The results showed that, the total length of Fiber2 gene of GZ-QL strain was 1 440 bp, with a 100.0% nucleotide homology of Fiber2 gene compared with Fiber2 gene of SCnj1601 strong strain isolated from Sichuan province and Fiber2 gene of NVD2 strong strain isolated from Beijing City. Besides, the nucleotide homology of Fiber2 gene between GZ-QL strain and FAdV-4 classical ON1 strain was 95.9%, and the two strains were located in the same evolutionary branch. There were 33 amino acid mutation sites in sequence of GZ-QL Fiber2 protein, and five amino acid were inserted into positions between the 11th and the 15th amino acids. The average hydrophilicity value of GZ-QL Fiber2 protein was-0.031 and its secondary structure was dominated by random coil(accounted for 51.8%), but there was no transmembrane domain or signal peptide, and the epitope distribution was uniform, which was predicted to have eight dominant epitope regions and concentrated in the N-terminal. The above results showed that, the GZ-QL Fiber2 protein of FAdV-4 Guizhou strain is relatively conserved and has good antigenicity in fowl adenovirus serotype 4, which has the potential to be the dominant antigen.

关 键 词：禽腺病毒血清4型 Fiber2基因 生物信息学分析 

分 类 号：Q785[生物学—分子生物学]