摘要
根据GenBank中猪圆环病毒2型(PCV2)全基因序列设计引物,从疑似患断奶仔猪多系统衰竭综合症(PMWS)的仔猪组织病料中分别扩增出PCV2的全基因,将全基因克隆并测序。根据PCV2 ORF2序列设计引物,扩增HN01株ORF2基因序列,克隆入真核表达载体pEGFP-N1。通过脂质体介导法转染细胞,观察表达产物的荧光情况。结果表明,成功从疑似PMWS病料中扩增出PCV2全基因序列,三者间同源性很高,与GenBank中登录的中国分离株核苷酸序列同源性较高,而与国外分离株的同源性较低。所构建的真核质粒pEGFP-N1-ORF2结构正确,能够在PK-15细胞中表达。
The full length gene was amplified from tissues of piglets suffered from PMWS （postweaning multisystemic wasting syndrome） , using the primers designed according to the whole genome published in GenBank. The resultant PCR products were cloned and sequenced. The ORF2 gene of HN01 strain was amplified and cloned into pEGFP-NI vector. The recombinant vector was transfecfioned by Lipofectamine. The results in- dicated three full length genes obtained by PCR had high homologies among and with the Chinese isolates in GenBank, while had low homologies between the foreign isolates. The eukaryotic plasmid pEGFP-N1-ORF2 which had the correct structure could be expressed in PK-15 cells.
出处
《安徽农业大学学报》
CAS
CSCD
北大核心
2011年第5期688-693,共6页
Journal of Anhui Agricultural University
基金
院博士科研启动基金(起止时间2008-2011)资助
关键词
猪2型圆环病毒
全基因
ORF2基因
序列分析
表达
porcine circovirus type 2
full length gene
ORF2 gene
sequence analysis
expression
