畜牧与兽医2012，Vol.44Issue(1)：8-11,4.
抗猪繁殖与呼吸综合征病毒转基因细胞系的建立
Establishment of a transgenic cell line with resistance to porcine reproductive and respiratory syndrome virus
摘要
Abstract
The objective of this study is to construct a transgenic cell line resistant to porcine reproductive and respiratory syndrome virus (PRRSV) infection. The lipid method was used to introduce the anti-PRRSV shRNA plasmid, pEGFP-Nl-shRNA, into PK-15 cells. After screened with G418, the fluorecent cells were separated and cultured repeatedly, and therefore an anti-PRRSV transgenic PK-15 cell line was established. The transfection methods and conditions were further optimized, and the constructed cell line was detected by PCR after frozen-thawed. The results showed that the optimal concentration of G418 was 600μg/mL, the transfection ratio of liposome to plasmid was 7:2, and the transfection time was 24 h. This study established successfully an anti-PRRSV transgenic PK-15 cell line, and thus applies a basis for function validation of anti-disease gene and somatic cell nuclear transplantation.关键词
猪繁殖与呼吸综合征病毒/PK-15细胞/转基因细胞系Key words
porcine reproductive and respiratory syndrome virus (PRRSV) / PK-15 cells/ transgenic cell line分类
农业科技引用本文复制引用
马汝钧,罗碧平,剧世强,芮荣..抗猪繁殖与呼吸综合征病毒转基因细胞系的建立[J].畜牧与兽医,2012,44(1):8-11,4.基金项目
博士学科点专项科研基金(20100097110008) （20100097110008）
国家转基因生物新品种培育重大专项(2009ZX08009-143B). （2009ZX08009-143B）
