鹅IL-2成熟基因的表达及重组蛋白生物活性检测  

Expression of GoIL-2 mature gene in E.coli

作　　者：卫娜[1] 鞠环宇[1] 王君伟[1] 

机构地区：[1]东北农业大学预防兽医微生物与免疫教研室,黑龙江哈尔滨150030

出　　处：《中国预防兽医学报》2009年第6期466-471,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：黑龙江省留学回国基金项目(No.LC02C08)

摘　　要：从PHA活化的东北白鹅外周血淋巴细胞中分离提取总RNA,利用反转录PCR技术扩增获得鹅IL-2(GoIL-2)基因,连接至克隆载体后进行测序鉴定,结果与预期大小一致。将去除信号肽的成熟基因亚克隆至原核表达载体,构建原核重组表达质粒并转化至大肠杆菌中进行诱导表达,用表达纯化的融合蛋白制备多克隆抗血清。此外,体外淋巴细胞增殖实验结果显示,鹅GoIL-2重组蛋白具有促进淋巴细胞增殖的活性,这为进一步研究GoIL-2的功能奠定了基础。The goose interleukin-2 （GolL-2） cDNA was amplified by reverse transcription-polymerase chain reaction （RT- PCR） from peripheral blood lymphocyte of Northeast Goose that pre-treated with PHA. The PCR product was inserted into the cloning vector and confirmed by sequencing. The mature gene with signal peptide removed was subcloned to expression vector of procaryon, transformed into Escberichia eoli for expression. The expressed protein was purified and used to prepare polyclonal antiserum. In addition, the lymphopoiesis experiment in vitro showed that the rGolL-2 had the ability to promote lymphopoiesis.

关 键 词：白细胞介素-2 克隆 表达 多克隆抗体 生物活性 

分 类 号：Q786[生物学—分子生物学]