抗蓝耳病shRNA转基因表达载体的构建与功能验证  

Construction and functional verification of anti-PRRS shRNA transgenic expression vector

作　　者：罗碧平[1,2] 马汝钧[1] 剧世强[1] 芮荣[1] 

机构地区：[1]南京农业大学动物医学院,南京210095 [2]金陵科技学院动物科学与技术学院,南京210038

出　　处：《上海农业学报》2013年第2期9-13,共5页Acta Agriculturae Shanghai

基　　金：博士学科点专项科研基金(20100097110008);江苏省研究生科研创新计划项目(CXZZ11-0682);江苏高校优势学科建设工程项目资助

摘　　要：将靶向猪繁殖与呼吸综合征病毒(PRRSV)的shRNA表达盒插入pEGFP-N1的EcoO109I酶切位点构建重组转基因载体pEGFP-G1,并在转基因Marc-145细胞验证其抑制PRRSV复制的功能。用pEGFP-G1转染Marc-145细胞24 h后感染欧洲型PRRSV,病毒感染后每天观察细胞病变(CPE)情况。试验结果表明:所获pEGFP-G1可明显抑制PRRSV复制。The PRRSV-targeting shRNA expression box was inserted into the EcoOl09I site of pEGFP-N1 to construct the recombinant transgenic vector pEGFP-G1 and its function of inhibiting PRRSV replication was verified in the transgenic Marc-145 cells. The Marc-145 cells were infected with Europe-type PRRSV at 24 h after transfected with pEGFP-NI,and the cytopathic effect was ob- served daily. The result showed that as compared with the control the pEGFP-G1 in the MARC-145 cells could significantly inhibit PRRSV replication.

关 键 词：猪蓝耳病 猪繁殖与呼吸综合征病毒 shRNA质粒构建 细胞病变效应 

分 类 号：S852.65[农业科学—基础兽医学]