摘要
为原核表达番鸭白细胞介素2(MdIL-2),本研究利用ConA刺激诱导番鸭脾淋巴细胞,采用RT-PCR技术扩增MdIL-2基因的编码序列,并将其克隆至pGEX-6P-1中,构建重组质粒pGEX-MdIL-2,转化E.coli BL21受体菌。测序结果表明,MdIL-2基因ORF全长420 bp,编码140个氨基酸。重组菌经IPTG诱导表达后,SDS-PAGE分析显示,重组蛋白约为40 ku。Western blot分析表明,重组蛋白能够被抗GST单克隆抗体特异性识别。MdIL-2基因的克隆及其原核表达为进一步进行MdIL-2的活性检测以及应用研究奠定了基础。
To express the Muscovy duck interleukin-2 （MdlL-2） gene, the encoding sequence of IL-2 was amplified by RT-PCR from Muscovy duck spleen lymphocytes stimulated with concanavalin A, and the 420 bp ORF of MdlL-2 was inscrtcd into pGEX-6P to constructed recombinant expression plasmid pGEX-MdlL-2 and expressed in F. coil by IPTG induction.SDS-PAGE analysis showed that the expressed product of the GST- MdlL-2 was 40 ku which was recognized by GST monoclonat antibody. The results could be used for further study of the bioactivity and application of MdlL2.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2012年第2期146-148,共3页
Chinese Journal of Preventive Veterinary Medicine
基金
福建省自然科学基金项目(Z0516012)
关键词
番鸭
白细胞介素2
克隆
表达
muscovy duck
interleukin-2 gene
cloning
expression
