摘要
试验针对J亚群禽白血病病毒的env及LTR基因,根据它们的保守序列,分别设计合成了4～5对siRNA,并将其克隆至pSilencer 4.1,构建siRNA重组表达质粒。将该质粒转染DF-1细胞6 h后以103TCID50接种ALV-J,利用Real-time RT-PCR在mRNA表达水平检测各重组质粒对病毒复制的影响。结果表明,与阴性对照相比,pSi4.1-env各重组质粒对env基因的抑制效果达到18.15%～63.43%,pSi4.1-LTR各重组质粒的抑制效果则达到19.37%～45.41%。
According to the env and LTR gene of avian leukosis virus subgroup J,4 and 5 short hairpin RNAs（shRNAs） targeting their conserved sequences were designed systhessised and cloned into pSilencer 4.1 vector respectively.The recombinant plasmids were transfected into DF-1 cell and infected with 103 TCID50 ALV-J at 6 hours post-transfection,the efficiency of RNA interfering was assayed by Real-time RT-PCR.The results revealed that the recombinant plasmids of pSi4.1-env inhibited virus growth by 18.15% to 63.43% and pSi4.1-LTR by 19.37% to 45.41% compared to negative control.
出处
《中国畜牧兽医》
CAS
北大核心
2012年第8期16-19,共4页
China Animal Husbandry & Veterinary Medicine
基金
NSFC-广东联合基金(U0831002)
国家肉鸡产业技术体系项目(nycytx-42-G3-03)
广东省自然科学基金项目(S2011010001946
10451064201005432)
