牛病毒性腹泻病毒HB-DCZ株E0基因的克隆与抗原表位预测  

Cloning and sequence analysis of E0 gene of bovine viral diarrhea virus HB-DCZ strain

作　　者：蒋禄峰[1] 赵月兰[2] 李苏楠[3] 常丽云[1] 张梦茜[1] 包永占[1] 秦建华[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071001 [2]河北农业大学动物医学院,河北保定071001 [3]吉林大学畜牧兽医学院,吉林长春130062

出　　处：《中国兽医学报》2013年第7期979-984,共6页Chinese Journal of Veterinary Science

摘　　要：根据GenBank上发表的牛病毒性腹泻病毒(BVDV)Oregon C24V株E0基因的核苷酸序列设计1对特异性引物,应用PCR技术扩增BVDV HB-DCZ株E0基因。将PCR产物克隆到pMD18-T载体,克隆产物进行序列测定与分析。结果显示,HB-DCZ cDNA体外扩增获得特异性条带,约为687bp。序列测定结果表明,HB-DCZ株E0基因由681个核苷酸组成,编码227个氨基酸。与已公开发表的BVDV其他毒株核苷酸和推导氨基酸序列同源性相似顺序依次为:CCSYD 99%和98.7%、QHZK10 98.4%和97.8%、VEDEVAC 98.2%和97.8%、Oregon C24V80.9%和89.9%、Yak 74.2%和81.1%。HB-DCZ与CCSYD、QHZK10以及VEDEVAC株的遗传距离较近,与国际标准毒株C24V、Yak株的遗传距离较远。运用Kyte-Doolittle方案预测氨基酸的亲水性,采用Karplus-Schulz方案预测蛋白质的柔性区域,按Jameson-Wolf方案预测抗原指数,利用Emini方案预测蛋白质的表面可及性。对预测结果综合分析,推测最有可能的B细胞表位位于N-端6～17、23～29、47～53、59～68、70～81、97～109、114～122、127～134、137～143、165～172、186～198和213～220。A pair of specific primers was designed on the Oregon C24V strain E0 nucleotide se- quence of bovine viral diarrhea virus published in Genbank and HB-DCZ strain E0 gene was am- plified by polymerase chain reaction（PCR）. The PCR product was cloned into pMD18-T vector, cloned products were sequenced and analyzed. Specific bands about the size of 687 bp was ampli- fied. Sequenced results showed that the E0 gene of HB-DCZ strains was composed of 681 nucleo- tides and encoding 227 amino acids. Compared with the other strains BVDV nucleotide and de- duced amino acid sequence homology was CCSYD 99% and 98.7% ,QHZK10 98.4X and 97.8%, VEDEVAC 98.2% and 97.8%,Oregon C24V 80.9% and 89.9% and Yak 74.2% and 81. IX,re- spectively. The genetic distance of HB-DCZ strain is close to CCSYD, QHZK10,and VEDEVAC strain, far from Oregon C24V with international standards stain C24V and the Yak strain. The hy- drophilicity plot of E0 was predicted by the Kyte-Doolittle based on E0 sequence and the flexibil- ity plot, the antigenic index and the surface probability plot were by the methods of Karplus- Schulz,Jameson-Wolf and Emini, respectively. The B cell epitopes of E0 by these predicted results was analyzed which are most likely localized in or adjacent to its N-teminal No. 6-17,23-29,47-53, 59-68,70-81,97-109,114-122,127-134,137-143,165-172,186-198 and 213-220. Key words：bovine viral diarrhea virus;the E0 gene; cloning;sequence analysis;predicted of the antigenic epitope

关 键 词：牛病毒性腹泻病毒 E0基因 克隆 序列分析 抗原表位预测 

分 类 号：S852.653[农业科学—基础兽医学]