新城疫病毒Anhinga株HN基因的克隆与生物信息学分析  

Cloning and Bioinformatics Analysis of HN Gene of Newcastle Disease Virus Anhinga Strain

作　　者：何金娇[1,2] 毛雪飞 仇书兴[2] 李瑞 李鹏 董春秀[1] 刘洋洋 栾淼[1] 吴云舟 HE Jin-jiao;MAO Xue-fei;QIU Shu-xing;LI Rui;LI Peng;DONG Chun-xiu;LIU Yang-yang;LUAN miao;WU Yun-zhou(College of Life Sciences,Northeast Agricultural University,Harbin,Heilongjiang,150030,China;School of Life Sciences&Basic Medicine,Xinxiang University,Xinxiang,Henan,453003,China)

机构地区：[1]东北农业大学生命科学学院,黑龙江哈尔滨150030 [2]新乡学院生命科学与基础医学学院,河南新乡453003

出　　处：《动物医学进展》2022年第2期7-13,共7页Progress In Veterinary Medicine

基　　金：国家自然科学基金青年项目(31502050);东北农业大学“学术骨干”项目(19XG17);河南省高等学校重点科研项目(20A180026);河南省科技攻关计划项目(182102310091);新乡学院博士科研启动项目(1366020118)。

摘　　要：克隆新城疫病毒中等毒力株Anhinga(Anh)的血凝素-神经氨酸酶(HN)基因,用生物信息学软件分析其编码蛋白结构与功能的差异。根据已知的HN基因序列,通过Primer Premier 5.0设计引物,提取新城疫病毒总RNA,反转录成cDNA,PCR扩增HN基因,并将其连接到T载体上进行测序比对;用DNAman、DNAstar、MEGA7.0及在线分析预测软件对HN基因编码的蛋白进行生物信息学分析。结果显示,HN基因全长为1716 bp,与预期基因序列同源性达99.77%。其基本理化性质为亲水性跨膜蛋白,无信号肽序列,二级结构主要以无规则卷曲为主,属于神经氨酸酶CL21531超家族,三级结构与二级结构预测结果基本吻合,该蛋白存在多个潜在的B细胞表位。论文成功克隆并分析了HN基因所编码蛋白的主要特性及可能存在的抗原位点,为深入探究HN蛋白在NDV感染、增殖及抗肿瘤方面的作用奠定了基础。The objective was to clone the hemagglutinin neuraminidase(HN)gene of the mesogenic strain of Newcastle disease virus Anhinga(Anh)stain,and analyze the difference of the structure and function of the encoded protein by bioinformatics software.Based on the known HN gene sequence,primers were designed by primer premier 5.0 software.The total RNA of Newcastle disease virus was extracted and reversely transcribed into cDNA.The HN gene was amplified by PCR and connected toT vector for sequencing.The protein encoded by HN gene was analyzed by DNAMAN,DNAStar,MEGA7.0 and some online analysis and prediction software.The results showed that the total length of HN gene was 1716bp,and the homology with expected gene sequence was 99.77%.Its basic physical and chemical properties are hydrophilic transmembrane protein,no signal peptide sequence,and its secondary structure is mainly irregular coil,which belongs to neuraminidase CL21531 superfamily.The prediction results of tertiary structure are basically consistent with those of secondary structure.several potential B cell epitopes are present in this protein.In this study,we successfully cloned HN gene and analyzed the main characteristics and possible antigenic sites of the protein encoded by HN gene,which will provide a theoretical basis for further exploring the role of HN protein in NDV infection,proliferation and anti-tumor.

关 键 词：新城疫病毒 血凝素-神经氨酸酶蛋白 生物信息学 理化性质 

分 类 号：S852.659.5[农业科学—基础兽医学]