摘要
利用反转录-聚合酶链式反应（RT-PCR）技术及RACE方法扩增得到鸭肝炎病毒（DHV）浙江分离株Z10的全基因（5′,3′末端序列用RACE法扩增）及4株DHV分离株的VP1基因。结果表明,分离株Z10的全基因片段长7 689 bp,有1个大的开放读码框（ORF）,ORF位于626~7 326位核苷酸,编码2 249个氨基酸。Z10分离株全基因序列与GenBank登录的6株具有代表性的DHV核苷酸序列比对,同源性94.5%~98.4%;所测得的DHV分离株的VP1基因的序列与目前GenBank上发表的具有代表性的DHV-ⅠVP1基因进行比对分析,结果4株Ⅰ型DHV的VP1基因cDNA长度均为714 bp,编码238个氨基酸。4株DHV-Ⅰ之间VP1基因的核苷酸序列同源性为93%~99.7%,氨基酸序列同源性为95.0%~100%;与参考毒株VP1基因的核苷酸序列同源性为92.2%~100%,氨基酸序列同源性为95.0%~100%;表明各分离毒株的亲缘关系较近,属于同一基因群。
The complete genome of DHV-I Z10 isolate and complete VP1 gene of four isolates were amplified with RT-PCR method and rapid amplification of cDNA Ends（RACE） method（The cDNA fragments from the 3′ end and 5′ end of the viral genome were amplified using RACE method）.With the molecular biology soft ware,the genome of Z10 isolate was analyzed.The results were as follows：The DHV-I Z10 genome consisted of 7 689 nucleotides（excluding the poly（A） tail） and contained a single open reading frame（ORF）.Sequence analyses with six other DHV complete nucleotide sequences published in GenBank showed 94.5%-98.4% nucleotide identities.Sequence analyses revealed that the VP1 genes of these DHV-I isolates all had the same size of 714 most other reported DHV-I isolates and encode 238 amino acids compared with by as those of.Among the cited DHV-I isolates,they shared 92.2%-100% nucleotide and 95.0%-100% amino acid sequence identities in the VP1 gene.The analysis showed that the close genetic relationship strains belong to the same gene cluster.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2010年第8期1052-1055,共4页
Chinese Journal of Veterinary Science
基金
长三角重点资助项目(2005E60014)
浙江省自然科学基金资助项目(Y3080158)
