Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine embryos in vitro

doi:10.15302/J-FASE-2018229

Frontiers of Agricultural Science and Engineering

2019, Vol. 6

Issue (1): 66-72 https://doi.org/10.15302/J-FASE-2018229

本期目录

Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine embryos in vitro

Jianwen CHEN¹, Kaiyuan PAN¹, Zhen CHEN¹, Biao DING¹, Dandan SONG¹, Wenbin BAO²(

), Yunhai ZHANG¹(

)

¹. Anhui Provincial Laboratory of Local Livestock and Poultry, Genetical Resource Conservation and Breeding, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China
². College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China

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Abstract：

Porcine viral diarrhea is an acute and highly contagious enteric disease of pigs that causes huge economic losses worldwide. Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) are the main pathogens responsible for piglet viral diarrhea. However, currently there is no specific drug available for the effective treatment of viral diarrhea. Therefore, it is necessary to seek an effective method to diminish PEDV and TGEV infection rates. RNA interference has been applied successfully to inhibit the virus replication. It provides a potential strategy for breeding resistant pigs. In this study, four promoters and four short hairpin RNA (shRNA) vectors with LoxP sites at each end of the selectable marker genes were constructed to target PEDV and TGEV. These vectors were then transfected into porcine fetal fibroblasts, G418 resistant transfectants were confirmed by PCR and transgenic SCNT porcine blastocysts were obtained. These results have paved the way for future production of marker-free transgenic resistant to PEDV and TEGV pigs by SCNT.

Key words： piglet diarrhea RNAi PEDV TGEV transgenic resistance breeding

收稿日期: 2017-12-24 出版日期: 2019-02-25

Corresponding Author(s): Wenbin BAO,Yunhai ZHANG

引用本文:

. [J]. Frontiers of Agricultural Science and Engineering, 2019, 6(1): 66-72.
Jianwen CHEN, Kaiyuan PAN, Zhen CHEN, Biao DING, Dandan SONG, Wenbin BAO, Yunhai ZHANG. Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine embryos in vitro. Front. Agr. Sci. Eng. , 2019, 6(1): 66-72.

链接本文:

https://academic.hep.com.cn/fase/CN/10.15302/J-FASE-2018229
https://academic.hep.com.cn/fase/CN/Y2019/V6/I1/66

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