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通过计算机分析设计,人工合成了寡核苷酸引物,并优化了翻译起始区,应用PCR及重组DNA技术,获得了重组人白细胞介素6(IL—6)高效表达工程菌。从全菌SDS—PAGE考马斯亮蓝染色后薄层密度扫描分析表达产率为38.6％,分子量为2.1×10~4。以IL—6依赖性的小鼠杂交瘤细胞株B9用MTT法测定表达重组人IL—6的HGF活性为2×10~7U/L菌液。不同菌株表达重组人IL—6研究表明,以E.coli DH5α/pBV IL—6表达产率最高。
Through computer analysis and design, oligonucleotide primers were synthesized and the translation initiation region was optimized. Recombinant human interleukin-6 (IL-6) highly expressing engineering bacteria was obtained by PCR and recombinant DNA technology. The whole cell SDS-PAGE Coomassie brilliant blue stained by TLC scanning, the expression yield was 38.6% and the molecular weight was 2.1 × 10 ~ 4. The HGF activity of the recombinant human IL-6 was determined to be 2 × 10 -7 U / L using the MTT assay in an IL-6-dependent mouse hybridoma cell line B9. Different strains of recombinant human IL-6 studies have shown that the highest expression rate of E. coli DH5α / pBV IL-6.