摘要:丝氨酸蛋白酶家族活性中心由一组特征性氨基酸残基构成，其中有一个是丝氨酸。该蛋白酶家族的作用是断裂大分子蛋白质中的肽键，使之成为小分子蛋白质。丝氨酸蛋白酶家族成员执行多种生理功能，具有广泛的研究与应用价值。本论文利用基因工程技术，构建P.ananatis丝氨酸蛋白酶原核表达载体，在大肠杆菌中表达重组丝氨酸蛋白酶。重组质粒的酶切鉴定和重组菌诱导表达后的蛋白电泳结果证明了本研究成功构建出重组丝氨酸蛋白酶原核表达载体。本研究为新型丝氨酸蛋白酶进一步的开发利用奠定了基础。

关键词 丝氨酸蛋白酶；原核表达载体；P.ananatis

Abstract:Activity center of serine protease family is constituted by a set of characteristic amino acid residues, which has a serine. The main function of this protease family is to break macro- molecular proteins to smaller ones in peptide bond. Serine protease family members perform a variety of physiological functions and have a wide range of research and application value. In the paper, a prokaryotic P. ananatis serine protease vector was constructed with genetic engineering technology, and the recombinant serine protease was expressed in E. coli. Restriction endo- nuclease analysis of recombinatant plasmid and electrophoresis of induced protein from recombinant bacteria show the successful construction of the recombinant serine protease prokaryotic expression vector. The paper provides foundation for the further development and utilization of novel serine protease.

Keywords  Serine protease  Prokaryotic expression vector  P. ananatis