稳定高效表达猪CD163的Marc-145细胞系的构建与鉴定

doi:10.16431/j.cnki.1671-7236.2015.08.022

›› 2015, Vol. 42 ›› Issue (8): 2074-2080.doi: 10.16431/j.cnki.1671-7236.2015.08.022

Establishment and Identification of One Marc-145 Cell Line Stably and Highly Expressing Porcine CD163

WU Xiang-ju^1,2, LI Fang-tao¹, CHEN Lei^2,3, CONG Xiao-yan^2,3, SUN Wen-bo^2,3, YU Jiang^2,3, CHEN Zhi^2,3, GUO Li-hui^2,3, WU Jia-qiang^2,3, DU Yi-jun^2,3, ZHAO Xiao-min¹, WANG Jin-bao²

1. Key Laboratory of Animal Biotechnology and Disease Control and Prevention of Shandong Province, College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an 271000, China;
2. Shandong Key Laboratory of Animal Disease Control and Breeding, Jinan 250100, China;
3. Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan 250100, China

Received:2015-01-21 Online:2015-08-20 Published:2015-08-27

Abstract

Abstract: This study was attempted to generate one Marc-145 cell line stably and highly expressing porcine CD163 (pCD163) and set the foundation for PRRSV isolation and vaccine production.CD163 was shown to be a cellular receptor capable of mediating infection of PRRSV non-permissive cell lines.The pCD163 gene was amplified by RT-PCR from porcine alveolar macrophages and cloned into the eukaryotic expression vector pCI-neo, then the positive plasmid pCI-pCD163 was transfected into Marc-145 cells.After selecting with G418 and subcloning for 3 times, Marc-145 cell line expressing pCD163 was established.IFA results indicated that the fluorescence of pCD163-Marc cells was significantly brighter than Marc-145 cells;Western blotting results indicated that the pCD163-Marc cells could express higher levels of CD163 and the expression level was 8.7 times higher than Marc-145 cells.The pCD163-Marc cell line could be stably passaged for 20 passages and the expression level of CD163 was similar with different passages, which would be a valuable tool for facilitating virus propagation and vaccine production.

Key words: Marc-145; CD163; stable cell line

CLC Number:

Q813

WU Xiang-ju, LI Fang-tao, CHEN Lei, CONG Xiao-yan, SUN Wen-bo, YU Jiang, CHEN Zhi, GUO Li-hui, WU Jia-qiang, DU Yi-jun, ZHAO Xiao-min, WANG Jin-bao. Establishment and Identification of One Marc-145 Cell Line Stably and Highly Expressing Porcine CD163[J]. , 2015, 42(8): 2074-2080.

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Establishment and Identification of One Marc-145 Cell Line Stably and Highly Expressing Porcine CD163

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