携Hsp70的PRRSV ORF5与ORF6基因重组质粒构建及在Cos-7细胞中的表达

›› 2011, Vol. 23 ›› Issue (3): 0-499.

• 动物科学 •

携Hsp70的PRRSV ORF5与ORF6基因重组质粒构建及在Cos-7细胞中的表达

左兰1,颜其贵1,2,*,刘彬3,李双江3

1四川农业大学动物生物技术中心,四川雅安 625014；2四川农业大学动物疫病与人类健康四川省重点实验室,四川雅安 625014；3四川农业大学动物医学院,四川雅安 625014

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-05-25 发布日期:2011-05-25

Construction of eukaryotic recombinant expression vector containing Hsp70 and PRRSV ORF5, ORF6 genes and its responses in Cos-7 cell line

ZUO Lan;YAN Qi-gui;*;LIU Bin;LI Shuang-jiang

1Animal Biotechnology Centre, Sichuan Agricultural University, Yaan 625014, China; 2Key Laboratory of Animal Disease and Human Health of Sichuan Province, Sichuan Agricultural University, Yaan 625014, China;3College of Animal Medical, Sichuan Agricultural University, Yaan 625014, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-05-25 Published:2011-05-25

摘要/Abstract

摘要： 为构建高效猪繁殖与呼吸综合征病毒(PRRS)DNA疫苗,利用Hsp70作为免疫佐剂增强免疫功能的特点,构建了含PRRSV ORF5,ORF6基因的融合表达载体pCI-ORF5-ORF6A,以及携带HSP70基因的融合表达载体pCI-ORF5-ORF6B-Hsp70,转染Cos-7细胞。IFA检测结果显示,两个融合表达载体均能在Cos-7细胞中表达；Western blot检测证实转染质粒pCI-ORF5-ORF6A和 pCI-ORF5-ORF6B-Hsp70在42.7 Ku,112.2 Ku分别表达出特异性条带。结果显示所构建的重组质粒能在Cos-7细胞内进行良好的表达,且表达的蛋白具有免疫原性。

关键词: PRRSV, Hsp70, Cos-7细胞

Abstract: In order to construct efficient porcine reproductive and respiratory syndrome (PRRS) DNA vaccines and carrying HSP70 gene which can be used as an adjuvant enhancing immune function, we constructed the fusion expression vector PCI-ORF5-ORF6A and PCI-ORF5-ORF6B-HSP70 which comprehended PRRSV ORF5 and ORF6 genes. The two kinds of fusion expression vectors were transfected Cos-7 cells. IFA results showed that the two fusion expression vector in Cos-7 cells could express and western blot test confirmed that the specific bands about 42.7 Ku and 112.2 Ku could be found from transfection of plasmid pCI-ORF5-ORF6A and pCI-ORF5-ORF6B-Hsp70. It showed that the recombinant plasmid in Cos-7 cells were efficiently transcribed and expressed, and the expression of the protein was immunogenic．

Key words: PRRSV, Hsp70, Cos-7 cell

左兰;颜其贵;*;刘彬;李双江. 携Hsp70的PRRSV ORF5与ORF6基因重组质粒构建及在Cos-7细胞中的表达[J]. , 2011, 23(3): 0-499.

ZUO Lan;YAN Qi-gui;*;LIU Bin;LI Shuang-jiang . Construction of eukaryotic recombinant expression vector containing Hsp70 and PRRSV ORF5, ORF6 genes and its responses in Cos-7 cell line[J]. , 2011, 23(3): 0-499.

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携Hsp70的PRRSV ORF5与ORF6基因重组质粒构建及在Cos-7细胞中的表达

Construction of eukaryotic recombinant expression vector containing Hsp70 and PRRSV ORF5, ORF6 genes and its responses in Cos-7 cell line

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