CAJ | 학술논문

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BL21(DE3),并经IPTG诱导表达,表达量达到菌体总蛋白的20%,并具有良好的活性.结论:证明双顺反子系统可以有效增加bFGF 的表达量.
목적:유우인감성성섬유세포생장인자편마기인결구존재불리우원핵표체적인소,구건료쌍순반자적대장간균표체계통,이기제고기표체량.방법:기중질립표체재체pET-3c휴대적T7서균체Φ10기인N단안기산적편마서렬위제일개순반자,bFGF편마기인위제이개순반자,이자동처우T7계동자적하유.결과:장중조자전도표체균BL21(DE3),병경IPTG유도표체,표체량체도균체총단백적20%,병구유량호적활성.결론:증명쌍순반자계통가이유효증가bFGF 적표체량.
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.