畜牧兽医学报2012，Vol.43Issue(10)：1657-1663,7.
山羊白细胞介素-4成熟蛋白基因的原核表达及其生物学活性检测
Expression of Goat Mature Interleukin-4 Gene in vitro and Bioactivity Detection of Recombinant Protein
摘要
Abstract
The aim of the present study was to express goat mature interleukin-4 gene in E. coli and to test the bioactivity of the recombinant protein. The mature GIL-4 (mGIL-4) was amplified using pMD18-T-pGIL-4 (pGIL-4, goat IL-4 complete sequence) as template by PCR. The recom-binant plasmid of pET-32a( + ) containing mGIL-4 gene was transformed into E. coli BL21, and induced by IPTG. The recombinant protein expressed in E. coli was purified by modified method and refolded. Lymphocyte transformation test was performed to detect the bioactivity of recombi-nant protein to stimulate the proliferation of T lymphocytes in goat's peripheral blood mononu-clear cells (PBMC). Sequence analysis of mGIL-4 demonstrated an open reading frame (ORF) of 339 base pairs encoding for 112 amino acids. The recombinant protein was approximately 31. 3 kD and was mainly existed in the inclusion body. Through improved purification methods, highly purified recombinant protein was obtained. The bioactivity test proved that purified recombinant protein was able to significantly stimulate the proliferation of goat peripheral blood T lympho-blasts in a dose-dependent manner and the recombinant protein of 8 μg ? mL-1 presented the high-est stimulation. The data provided foundations for the scale production and the application as adjuvant of the goat recombinant interleukin-4.关键词
山羊/IL-4/表达/生物学活性Key words
goat/ IL-4/ expression/ bioactivity分类
农业科技引用本文复制引用
张晓燕,严若峰,徐立新,宋小凯,李祥瑞..山羊白细胞介素-4成熟蛋白基因的原核表达及其生物学活性检测[J].畜牧兽医学报,2012,43(10):1657-1663,7.基金项目
国家"863"项目(2011AA10A211) （2011AA10A211）
江苏高校优势学科建设工程资助项目 （）
