小熊猫IL-17的克隆及原核表达  

Cloning and Prokaryotic Expression of IL-17 of Red Panda

作　　者：赵玄多 徐素慧 杨雯昱[1] 高洋[1] 修云芳 陈玉村 陈德坤[1] 

机构地区：[1]西北农林科技大学动物医学院,陕西杨凌712100 [2]海峡(福州)大熊猫研究交流中心,福建福州350001

出　　处：《动物医学进展》2014年第6期40-44,共5页Progress In Veterinary Medicine

基　　金：横向合作项目(201303020704)

摘　　要：为制备小熊猫白介素17(IL-17)表达产物,提取活化的小熊猫外周血单个核细胞总RNA,合成cDNA,以此为模板克隆IL-17的成熟肽编码序列,将其重组到原核表达载体pET-32a,并将其转入到BL-21(DE3)中,进行序列分析。重组载体经IPTG诱导表达,表达产物经镍柱纯化。结果显示,小熊猫IL-17的CDS区由462bp组成,含23个氨基酸组成的信号肽,重组IL-17由130个氨基酸组成。SDS-PAGE显示,原核表达产物在诱导温度25℃,IPTG浓度0.5mmol/L,诱导时间20h的条件下,能够获得最佳表达。To obtain the expression products of Red panda IL-17,total RNA was extracted from activated PBMC of Red panda and cDNA was synthesized and used as template for PCR subsequently.Mature pep-tide region was cloned and inserted into the prokaryotic expression vector PET32a and the recombinant plasmid was transformed into the competent cell BL21 （DE3）and a series of identification,sequence analy-sis were taken.The expression products were induced by IPTG and purified by the method of Ni-NTA. The results of sequence analysis suggested that the CDS region of Red panda IL-17 comprised 462 bp and the recombinant protein was composed of 130 animo acids and 23 of which formed the signal peptide.The results of SDS-PAGE showed the optimal induction condition was culturing with a total concentration of 0.5mmol/L IPTG under 25 ℃ for 20 h.

关 键 词：小熊猫 白介素 17 基因克隆 原核表达 纯化 IL-17 

分 类 号：S852.2[农业科学—基础兽医学]