Establishment and Identification of One Marc-145 Cell Line Stably and Highly Expressing Porcine CD163 |
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Authors: | 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 |
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Institution: | 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 |
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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. |
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Keywords: | Marc-145 CD163 stable cell line |
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