C基因型牛副流感3型病毒分离株全基因组测序及分析  

Sequencing of complete genome of bovine parainfluenza 3 virus genotype C

作　　者：冉旭华[1] 张峣[1] 卢元赫 曹思[1] 张玲玲[1] 刘阳阳[1] 倪宏波[1] 闻晓波[1] 

机构地区：[1]黑龙江八一农垦大学预防兽医学省重点实验室,黑龙江大庆163319

出　　处：《中国生物制品学杂志》2016年第6期595-599,共5页Chinese Journal of Biologicals

基　　金："十二五"农村领域国家科技计划课题(2012BAD12B03-3);黑龙江省农垦总局攻关课题(HNK125B-11-08A);黑龙江八一农垦大学研究生创新科研项目(YJSCX2015-Y19);黑龙江省大学生创新创业训练计划项目(201510223020)

摘　　要：目的对宁夏地区分离得到的1株C基因型牛副流感3型病毒(bovine parainfluenza 3 virus,BPIV3)进行全基因组测序,并进行同源性及进化树分析。方法参考Gen Bank上登录的BPIV3中国分离株SD0835基因组序列设计5对引物,覆盖病毒基因组全长,提取BPIV3总RNA,以其为模板,进行RT-PCR扩增,扩增产物与p MD誖18-T Vector连接,连接产物转化大肠埃希菌DH5α感受态细胞,将阳性克隆进行核苷酸测序,测序结果通过分子生物学软件进行拼接,并与Gen Bank上登录的参考序列进行比较,构建系统进化树。结果成功分离得到1株BPIV3,命名为NX49,NX49株全基因组核酸序列为15 474 nt,提交至Gen Bank的序列号为KT071671。NX49与中国山东C型分离株SD0835的同源性最高,为99.3%;与中国内蒙古A型分离株NM09的同源性为82.5%;与澳大利亚B型分离株Q5592的同源性为81.4%。结论 NX49与SD0835同属一个进化分枝,但由于地域及时间上的不同,可能造成它们之间在基因水平上存在差异。Objective To sequence the complete genome of a bovine parainfluenza 3 virus（BPIV3） genotype C strain isolated from Ningxia Region in China, analyze the homology and construct a phylogenetic tree. Methods Five pairs of primers specific to full-length genome of BPIV3 were designed on the basis of the reference strain SD0835 isolated in China in Gen Bank. The total RNA of BPIV3 was extracted and used as a template for RT-PCR. The PCR product was cloned to p MD 18-T Vector, while the constructed recombinant plasmid was transformed to competent E. coli DH5αcells, and the selected positive clones were subjected to nucleotide sequencing. The sequencing results were spliced by molecular biological software and compared with the reference sequences in Gen Bank, based on which a phylogenetic tree was constructed. Results A BPIV3 strain was successfully isolated and named as NX49, of which the complete genome sequence consisted of 15 474 nt. The strain was submitted to Gen Bank, of which the accession No. was KT071671. The homologies of NX49 strain were 99. 3% to SD0835 strain of genotype C isolated in Shandong, China, 82. 5% to the NM09 strain of genotype A isolated in Inner Mongolia, China, and 81. 4% to Q5592 strain of genotype B isolated in Australia.Conclusion The NX49 strain was clustered with SD0835 strain. However, the diversities in geography and isolation time may cause differences between them at the genetic level.

关 键 词：牛副流感3型病毒 全基因组 C基因型 测序 

分 类 号：S852.653[农业科学—基础兽医学] Q755[农业科学—兽医学]