摘要
以RT-PCR法从鸡脾脏细胞中扩增出鸡白细胞介素2编码基因,通过双酶切、连接步骤克隆进原核表达载体pQE30及pGEX-4T-3,构建了鸡白细胞介素2基因的重组原核表达质粒pQE30-chIL-2及pGEX-chIL-2.重组质粒转化大肠杆菌JM109后经诱导表达,分别得到两种表达产物.以SDS-PAGE和Westernblot检测确认表达产物为带6组氨酸标签及GST蛋白的融合鸡白细胞介素2,分子量分别为16 kDa及39.5 kDa.表达蛋白与抗鸡白细胞介素2单抗均有良好的免疫结合性,进一步证实为鸡白细胞介素2.表达产物经过纯化复性后,具有明显促进鸡脾淋巴细胞增殖的作用.
The gene encoding chicken interleukin - 2 （chlL - 2） was amplified from chicken spleen ceils by RT - PCR. Following double digestion by restriction endonucleases and ligation, chlL - 2 was cloned into prokaryotic expression vectors of pQE30 and pGEX - 4T - 3 to construct recombinants of pQE30 - chlL - 2 and pGEX - chlL - 2, which were consequently transformed into E. coil JM109. Two kinds of recombinant protein were obtained with IPTG induction. The molecular weights of the expressed proteins were 16 kDa and 39.5 kDa on SDS - PAGE and they were regarded as 6His and GST labeled protein respectively as checked by Western blot. Both proteins revealed specific reaction to anti - chlL - 2 mAb, further confirming that recombinant proteins are chicken interleukin - 2. After purification and renaturation, the 6His- chlL- 2 showed biological activity in stimulating chicken lymphocyte proliferation.
出处
《福州大学学报（自然科学版）》
CAS
CSCD
北大核心
2006年第5期760-765,共6页
Journal of Fuzhou University(Natural Science Edition)
基金
福建省科技攻关计划重点项目(2003N083)
厦门市科技攻关计划重点项目(3502Z20031054)
集美大学科研启动基金(F03003)
