摘要
【目的】扩增口蹄疫病毒(Foot-and-mouth disease virus,FMDV)O/Akesu/58 CE39A株基因组3'末端序列,并对其核苷酸序列进行分析,为FMDV反向遗传学研究奠定基础。【方法】采用普通PCR法和3'RACE法扩增FMDV基因组3'末端序列,利用分子生物学软件对该序列进行分析。【结果】扩增了口蹄疫病毒基因组3'末端序列,3'UTR长98 nt,Poly(A)长度不同(19、23、24、25、27、29、43)。3'UTR序列二级结构含有2个茎环结构,分别位于8 101～8 140和8 146～8 190 nt。O/Akesu/58 CE39A株3'UTR序列与参考毒株O/MAY/3/2014(GenBank登录号:KY322672)和O/IND26(54)/2014(GenBank登录号:KJ825807)的核苷酸序列同源性最高,均为89.8%。【结论】普通PCR法和3'RACE法均可扩增出3'末端序列。同一毒株的亚克隆,其Poly(A)长度与基因扩增时所用方法有关。不同毒株的Poly(A)长度不同,可能是毒株本身属性,也有可能是由扩增方法不同造成。Poly(A)要么扩增成功后其长度不同,要么扩增失败。失败的原因可能是3'末端结构复杂,难于扩增。
【Objective】This project aims to amplify the 3'-end sequences of the O/Akesu/58 CE39A strain and conduct analysis of the nucleotide sequence in order to lay the foundation for the study of the reversed genetics of foot-and-mouth disease virus(FMDV).【Method】The 3'-end sequence of foot-and-mouth disease virus genome was amplified by general PCR and 3'RACE,and analyzed by molecular biology software.【Result】The 3'-end sequence of FMDV genome was successfully amplified and the length of 3'UTR was 98 nt,but the lengths of Poly(A)were different(19,23,24,25,27,29 and 43).The secondary structure of 3'UTR sequence contained two stem-loop structures,which were located in 8 101-8 140 nt and 8 146-8 190 nt,respectively.The 3'-end sequence of O/Akesu/58 CE39A showed the highest homologous with reported strain O/MAY/3/2014(GenBank accession no.KY322672)and O/IND26(54)/2014(GenBank accession no.KJ825807),and the similarity of these two reported strains with O/Akesu/58 CE39A was up to 89.8%.【Conclusion】The 3'-end sequences can be amplified by general PCR and 3'RACE.The length of Poly(A)of the same virus strain is related to the method used in gene amplification.The lengths of Poly(A)of different viruses are different,which may be attributed to the virus itself,or may be caused by different amplification methods.Poly(A)is either amplified or unsuccessful.The reason for the failure may be that the structure of the 3'-end is complex,so it is difficult to amplify.
作者
李金娜
苗书魁
马文戈
魏玉荣
汪萍
夏俊
陆桂丽
米晓云
沙依兰.卡依扎
王延
魏婕
黄炯
LI Jin-na;MIAO Shu-kui;MA Wen-ge;WEI Yu-rong;WANG Ping;XIA Jun;LU Gui-li;MI Xiao-yun;Shayilan Kayizha;WANG Yan;WEI Jie;HUANG Jiong(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China;Institute of Veterinary Medicine,Xinjiang Academy of Animal Sciences,Urumqi 830011,China)
出处
《新疆农业科学》
CAS
CSCD
北大核心
2018年第11期2150-2156,共7页
Xinjiang Agricultural Sciences
基金
国家重点研发计划项目"边境地区外来动物疫病阻断及防控体系研究"(2017YFD0501800)"边境地区家养动物外来动物疫病监测溯源技术研究"课题(2017YFD0501801)
新疆维吾尔自治区创新环境建设专项科技创新基地建设项目"新疆兽医微生物资源共享平台建设"(PT1809)
2014年新疆维吾尔自治区公益性科研院所基本科研业务经费资助项目(KY2014013)~~
