Yi Tianhong, Hong An, Xie Shanshan, Zhang Ling, Xie Qiuling, Dai Yun, Yu Rongjie
Bio-engineering Institute of Jinan University, Guangzhou 510632, China.
Sheng Wu Gong Cheng Xue Bao. 2008 Dec;24(12):2049-55.
To produce recombinant Maxadilan using gene engineering technology, the gene of recombinant Maxadilan which expressed in protocaryon were designed and synthesized according to the amino acid sequences of Maxadilan. The recombinant plasmid pKYB-MAX was constructed and transformed into host bacteria Escherichia coli strain ER2566. After the MAX-intein-CBD fusion protein was purified by chintin-affinity chromatography, the self-cleavage activity of the intein was induced by beta-mercaptoethanol and the recombinant Maxadilan was released from the chitin-bound intein tag. The molecular weight of peptides was determined by the laser flight mass spectrometry and the results was conformity with the theoretical value. The biological activity analysis showed that recombinant Maxadilan significantly enhanced the concentration of serum glucose.
利用基因工程技术生产重组麦克斯迪兰，根据麦克斯迪兰的氨基酸序列设计并合成了在原核生物中表达的重组麦克斯迪兰基因。构建重组质粒pKYB-MAX并将其转化到宿主菌大肠杆菌ER2566菌株中。通过几丁质亲和层析纯化MAX-内含肽-CBD融合蛋白后，用β-巯基乙醇诱导内含肽的自切割活性，使重组麦克斯迪兰从与几丁质结合的内含肽标签上释放出来。用激光飞行质谱法测定肽的分子量，结果与理论值相符。生物活性分析表明，重组麦克斯迪兰显著提高了血清葡萄糖浓度。
