重组人碱性成纤维细胞生长因子在大肠杆菌中的高效表达

HIGH EXPRESSION OF RECOMBINANT HUMAN BASIC FIBROBLAST GROWTH FACTOR IN ESCHERICHIA COLI

利用基因改造的方法可以优化外源基因在大肠杆菌中的表达。利用逆转录PCR技术从原代培养的人成纤维细胞中克隆出人碱性成纤维细胞生长因子基因 ,在不改变氨基酸序列的前提下对该基因的上游部分序列进行改造 ,并将其插入表达载体pET 3c ,转入大肠杆菌BL2 1 (DE3) ,IPTG诱导表达 ,表达蛋白占菌体总蛋白的 30 %以上 。

To improve the expression level of non fusion hbFGF in E coli , the coding sequence of human bFGF gene, which had been cloned from primarily cultured human fibroblast, was mutated according to the principle of lowering the GC content and increasing the codon preference After being ligated into pET 3c and transformed into BL21(DE3), the recombinant induced by IPTG Expression level was up to 30% of the total bacterial protein The result indicated that optimizing of the TIR would promote the expression level of recombinant protein