Cloning,Sequence Analysis and Prokaryotic Expression of Mink Interleukin-4 Gene |
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| Authors: | FAN Si-ning ZHANG Hai-ling ZHAO Jian-jun WANG Yang HU Bo LV Shuang MA Fan-shu LI Xin-tong LING Ming-yu ZHAO Hui LIAN Shi-zhen YAN Xi-jun |
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| Institution: | 1. College of Animal Science and Technology, Jilin Agricultural University, Changchun 130118, China;
2. Division of Infectious Diseases of Special Economic Animal, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun 130112, China |
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| Abstract: | In order to obtain high purity of interleukin 4 (IL-4) protein,the specific primers were designed and synthesized according to the mink IL-4 gene sequence in GenBank. The IL-4 gene was amplified by RT-PCR using total RNA of the lymphocyte isolated from the peripheral blood that induced by phytohemagglutinin (PHA). The length of IL-4 gene sequence was 399 bp which encoded 132 amino acids. Phylogenetic analysis revealed that the amino acid sequences homology between mink and ferret was 99.2%,90.0% with Ailuropoda melanoleuca and Canis familiaris. Prokaryotic expression vector pProEX-HTb-IL-4 was constructed and induced by IPTG. The recombinant protein of 15 ku was isolated by SDS-PAGE and detected by Western blotting. Highly purified recombinant protein of mink IL-4 was obtained by His-Trap HP affinity columns method. This research laid the foundation for the further studies on the biological function of mink IL-4 gene. |
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| Keywords: | mink interleukin-4 sequence analysis prokaryotic expression |
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