水貂白细胞介素-4基因的克隆、序列分析与原核表达

doi:10.16431/j.cnki.1671-7236.2017.01.004

Abstract

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.

Key words: mink; interleukin-4; sequence analysis; prokaryotic expression

CLC Number:

Q785

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. Cloning,Sequence Analysis and Prokaryotic Expression of Mink Interleukin-4 Gene[J]. , 2017, 44(1): 30-37.

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Cloning,Sequence Analysis and Prokaryotic Expression of Mink Interleukin-4 Gene

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