Cloning and Eukaryotic Expression Plasmid Construction of Porcine Interleukin-18 Gene |
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| Authors: | SHI Ang SHI Qing-he LI Xin-guo WANG Yu-guo YANG Li LI Shuang-shuang CHEN Lu WANG Chuan-qing |
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| Institution: | College of Animal Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002, China |
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| Abstract: | To obtain recombinant eukaryotic expression plasmid of porcine interleukin-18(IL-18), the whole gene of porcine IL-18 gene was amplified from porcine spleen, lung and lymph nodes by RT-PCR and cloned into eukaryotic expression vector pZJ-1. The recombinant expression plasmid pZJ-IL-18 were identified by enzyme digestion and sequencing analysis, and was transfected into 293T cells.The expression of IL-18 was detected by Real-time PCR and Western blotting in both gene and protein levels. The results showed that the eukaryotic expression plasmid of porcine IL-18 was constructed and could express transiently in 293T cells. Western blotting result confirmed that porcine IL-18 polyclonal antibody could react specifically with approximately 17 ku expression products,and indicated that IL-18 could express correctly and be responsive.This study constructed the eukaryotic expression plasmid of porcine IL-18 gene which could express transiently in 293T cells, and laid the foundation for studying function of IL-18. |
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| Keywords: | porcine IL-18 gene eukaryotic expression vector clone |
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