作　　者： ; ; ; ; ; ;

机构地区： 广东海洋大学水产学院广东省水产经济动物病原生物学及流行病学重点实验室

出　　处： 《广东海洋大学学报》 2008年第3期45-49,共5页

摘　　要： 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。 In order to further the study of the biological activity and function of ASP from Vibrio alginolyticus, the recombinant plasmid pET23d-asp were transformed into the host E. coli BL21（DE3）. The expressed product was purified and analyzed, and the expression conditions were optimized. The soluble ASP was purified when the chromatogram column was eluted by 100 mmol/L imidazole eluting buffer. The purified ASP had enzymatic activity and its relative molecular weight was 42 000. When the expression was induced with 1 mmol/L Isopropyl-β-D-thiogalactopyranoside （IPTG） for 16 h at 28 ℃, the maximum amounts of soluble proteins were obtained.

关 键 词： 溶藻弧菌 基因 基因构建 基因表达 表达条件 大肠杆菌

领　　域： [生物学] [轻工技术与工程] [轻工技术与工程]