溶藻弧菌asp基因在大肠杆菌中表达活性研究及条件优化Optimization of Expression Conditions and Activity of asp Gene from Vibrio alginolyticus in Escherichia coli
蔡双虎,鲁义善,吴灶和,简纪常,汤菊芬,黄郁葱
摘要(Abstract)：
为进一步研究溶藻弧菌碱性丝氨酸蛋白酶(ASP)蛋白生物学活性和功能,将构建的pET23d-asp在大肠杆菌BL21(DE3)中表达,对表达产物进行纯化分析,并优化表达条件。结果表明:在咪唑洗脱缓冲液浓度为100 mmol/L时,表达的可溶性ASP被大量洗脱,纯化的ASP相对分子质量约为42 000,具有蛋白活性;在诱导温度28℃、异丙基-β-D硫代半乳糖苷(Isopropyl-β-D-thiogalactopyranoside,IPTG)浓度1 mmol/L及诱导时间16 h时,表达的活性ASP蛋白最多。
关键词(KeyWords)： 溶藻弧菌;asp基因;基因构建;基因表达;表达条件;大肠杆菌
基金项目(Foundation): 广东省重大专项(2004A20401001);; 广东省重大科技兴海(兴渔)项目(204108)
作者(Author): 蔡双虎,鲁义善,吴灶和,简纪常,汤菊芬,黄郁葱
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