靶向ORFV-DNA ploymerase基因shRNA表达载体的构建  

Construction of ShRNA Recombination Expression Vector Targeting ORFV-DNA Polymerase

作　　者：于永忠[1] 郭雯[1] 吴欣媛[1] 王金珍[1] 崔玉东[1] 

机构地区：[1]黑龙江八一农垦大学生命科学技术学院,大庆163319

出　　处：《黑龙江八一农垦大学学报》2012年第4期38-41,共4页journal of heilongjiang bayi agricultural university

基　　金：国家自然科学基金面上项目(31172353);黑龙江省教育厅科学技术研究项目(11551322)

摘　　要：羊口疮(Orf)是由羊口疮病毒(ORFV)引起的人畜共患的一种急性、接触性和具有高度嗜上皮性传染病,主要感染绵羊、山羊和人。由于该病缺乏全身性感染症状,因此在防治上也缺乏有效的疫苗或抗体。RNA干扰技术是目前较为成熟的抑制目的基因表达生物技术。ORFV的DNA ploymerase是病毒复制的关键酶。研究运用RNA干扰技术针对ORFV的DNAploymerase基因进行体外基因沉默研究,通过网络自动筛选平台设计并合成3个short-hairpin RNAs(shRNAs)片段,并与含U6启动子的pLL3.7质粒构建重组载体,结果表明pLL3.7-D596为重组阳性,进一步测序结果正确。研究可以为靶向ORFV-DNAploymerase基因的体内基因沉默提供参考数据。Orf（contagious ecthyma,contagious pustular dermatitis,scabby mouth） was an acute skin zoonosis caused by orf virus（ORFV）,which could infect sheep,goats and humans.Because of the lack of systemic infection symptom,the prevention was also short of effective vaccines or antibodies.RNA interference technology was the relatively mature biotechnology which was used to inhibit the expression of target genes.DNA polymerase of ORFV played an important role as key enzyme related to replication.In this study,RNA interference technology was applied to vitro gene silencing research on DNA polymerase gene of ORFV,by automatic screening platform of network,synthesized 3 shRNAs fragments which with pLL3.7 plasmid contained U6 promoter to build recombination vector.The result showed that pLL3.7-D596 was positive clone followed by sequencing,which could provide reference data to vivo gene silencing of DNA polymerase of ORFV.

关 键 词：羊口疮病毒 DNA聚合酶 shRNA片段 重组载体 

分 类 号：S482[农业科学—农药学]