猪圆环病毒2型SD2分离株ORF3基因转录分析  

Transcription Analysis of ORF3 Gene in PCV2 Isolate SD2

作　　者：丁鹏[1,2] 李俊[2] 于周[1,2] 程凯慧[1,2] 吴家强[2] 王金宝[2] 

机构地区：[1]青岛农业大学,青岛266109 [2]山东省农业科学院畜牧兽医研究所,济南250100

出　　处：《中国畜牧兽医》2009年第8期128-130,共3页China Animal Husbandry & Veterinary Medicine

基　　金：山东省自然基金(Y2006D23);山东省农业科学院高技术自主创新基金(2007YCX017-04)

摘　　要：对PCV2 SD2株(DQ478947)进行了序列分析发现,其含有11个开放阅读框,与以往分离株开放阅读框有较大差异。根据GenBank发表的PCV2序列设计特异性引物,扩增ORF3基因,预计扩增片段大小为209 bp。分别在接毒后12、24、364、8 h,用Trizol法自病毒细胞裂解液提取总RNA,然后用DNase I处理,除去残留的病毒DNA,利用醋酸纤维素分离mR-NA,用上述引物进行RT-PCR扩增。在接毒24 h后扩增出的目的片段大小与预计扩增片段大小相符合,测序结果表明扩增片段与PCV2 SD2(DQ478947)ORF3基因序列相符。由此证实,PCV2山东分离株ORF3基因可在接毒24 h后于转录水平表达,为ORF3基因的深入研究打下了基础。The sequence analysis indicated that PCV2 SD2 （DQ478947） included 11 opening reading frames at least and some opening reading frames were different from other isolated strains. A pair of primers were designed according to the PCV2 sequence published on GenBank to amplify gene ORF3 of PCV2, the predict fragment size respectively was 209 bp. At 12, 24, 36, 48 hours post-PCV2 infection, total RNA was extracted from viral cell culture using Trizol reagent and treated with DNase I to eliminate the remained virus DNA. mRNA was separated by acetyl cellulose. ORF3 were amplified by RT-PCR. Results showed the amplified fragments size matched the estimate increase the fragment size totally, the sequence result indicated that the increased fragments and the corresponding sequences in PCV2 SD2 （DQ478947） were identical. Thus, the ORF3 gene of PCV2 SD2 isolate could express at the level of transcription after 24 hours post-infection.

关 键 词：猪圆环病毒2型 ORF3基因 转录分析 

分 类 号：S852.659.2[农业科学—基础兽医学]