摘要
为构建番鸭小鹅瘟病毒感染性克隆,进行水禽细小病毒基因组结构及功能研究,根据NCBI发表的GPV B株、MDPV FM株与MDGPV PT株核苷酸序列设计并合成了MDGPV特异性引物,进而应用PCR技术分4段扩增并克隆了覆盖MDGPV全长基因组的4个片段。经过拼接与测序,将MDGPV全长DNA定向克隆到质粒pBluescriptⅡKS(+)中,获得了MDGPV PT株全长基因组DNA克隆pSK-PT1234。与MDGPV亲本PT株及NCBI登录的其他水禽细小病毒代表株序列比对发现,该克隆在NS与VP编码区间隔处存在人为引入Bsp1407Ⅰ酶切位点。番鸭小鹅瘟病毒PT株全长DNA克隆成功构建,为下一步病毒的拯救奠定坚实的基础。
To develop a reverse genetics system of Muscovy duck-origin goose parvovirus（MDGPV）,4pairs of oligonucleotides were designed for PCR to 4overlapping fragments covering MDGPV PT genome based on the full-length genomic sequences of strains GPV B,MDPV FM and MDGPV PT.A MDGPV full-length DNA clone pSK-PT1234 was cloned to pBluescriptⅡKS（＋）and confirmed by plasmid PCR and sequencing.Comparing to the published sequences of waterfowl parvovirus strains,this clone possessed an introduced artificially restriction enzyme site Bsp1407Ⅰ located in the small noncoding gap of 18 nucleotides between the ORFs NS and VP.MDGPV strain PT full-length DNA clong has been produced successfully,which has laid a good foundation for the further rescued research.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2015年第7期1064-1068,共5页
Chinese Journal of Veterinary Science
基金
国家公益性行业(农业)科研专项(201003012)
国家自然科学基金委员会-福建省人民政府促进海峡两岸科技合作联合基金(U1305212)
福建省自然科学基金资助项目(2012J01110)
福建省农科院创新团队项目(STIF-Y02)
福建省农业科学院"青年科技英才百人计划"(YC2015-18)
国家青年科学基金资助项目(31402236)
关键词
番鸭小鹅瘟病毒
全长DNA克隆
感染性克隆
Muscovy duck-origin goose parvovirus（MDGPV）
full-length DNA
infectious DNA clone
