利用双顺反子系统在大肠杆菌中高表达bFGF(英文)  

The construction of dicistron system to improve the expression level of bFGF in Escherichia coli

作　　者：黄更生[1] 汪炬[2] 孙奋勇[2] 李志英[2] 张玲[2] 洪岸[2] 李贵生[1] 

机构地区：[1]暨南大学动物学实验室,广东广州510632 [2]暨南大学生物工程研究所,广东广州510632

出　　处：《中国病理生理杂志》2003年第5期627-631,共5页Chinese Journal of Pathophysiology

基　　金：TheNational86 3Plan (No .z18- 0 3- 2 9)

摘　　要：目的 :由于人碱性成纤维细胞生长因子编码基因结构存在不利于原核表达的因素 ,构建了双顺反子的大肠杆菌表达系统 ,以期提高其表达量。方法 :其中质粒表达载体pET - 3c携带的T7噬菌体Φ10基因N端氨基酸的编码序列为第一个顺反子 ,bFGF编码基因为第二个顺反子 ,二者同处于T7启动子的下游。结果 :将重组子转导表达菌BL2 1(DE3) ,并经IPTG诱导表达 ,表达量达到菌体总蛋白的 2 0 % ,并具有良好的活性。结论AIM: To investigate the condition necessary for high-level expression of basic fibroblast growth factor (hbFGF) in Escherichia coli, a dicistron system was constructed. METHODS: The phage T7 gene φ10 N-terminal sequence, present in the expression plasmid pET-3c, was incorporated into the first cistron, the coding region for bFGF was incorporated into the second, and both were under the control of a transcription promotor recognized by the bacteriophage T7 RNA polymerase. RESULTS: Recombinants were transformed into BL21(DE3) and induced by IPTG for expression, accumulation of up to 20% of the total cell protein of bFGF were obtained, showing bioactivity indistinguishable from standard protein. CONCLUSION: It suggested that such dicistron system was effective for enhancing hbFGF expression.

关 键 词：双顺反子 大肠杆菌 碱性成纤维细胞生长因子 质粒表达载体 pET—3c 

分 类 号：Q78[生物学—分子生物学] R363[医药卫生—病理学]