Lin Y, Lu S, Liang Z
Institute of Basic Medical Sciences, CAMS and PUMC, Beijing.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 1995 Apr;17(2):86-91.
Two kinds of expression plasmids carried Ref-Adaptor-(alpha-hANP) hybrid gene have been constructed. The adaptor GAA is the codon of glutamic acid which is a recognition site of the endoproteinase Glu-C. The expression of the heterologous fusion gene was controlled by the bacteriophage lambda PL promoter in the Escherichia coli expression system which is affected by the cIts857 repressor. These overproduced proteins were found in the form of inclusion bodies and constituted 35% and 48% of the total cellular protein respectively. The fusion protein was isolated and cleaved with Glu-C to liberate mature alpha-hANP, which demonstrated significant activities for reduction of blood pressure and vasorelaxation comparable to that of the chemically synthesized hormone. In this paper, the comparison between the two kinds of expression systems is also discussed.
已构建了两种携带Ref-衔接子-(α-人心房钠尿肽)杂合基因的表达质粒。衔接子GAA是谷氨酸的密码子，它是内蛋白酶Glu-C的识别位点。在受cIts857阻遏物影响的大肠杆菌表达系统中，异源融合基因的表达由噬菌体λ PL启动子控制。这些过量产生的蛋白质以包涵体的形式存在，分别占细胞总蛋白的35%和48%。分离融合蛋白并用Glu-C切割以释放成熟的α-人心房钠尿肽，其在降低血压和血管舒张方面表现出与化学合成激素相当的显著活性。本文还讨论了两种表达系统之间的比较。
