海马齿甜菜碱醛脱氢酶基因在大肠杆菌中表达

Expression of Recombinant Betaine Aldehyde Dehydrogenase of Sesuvium portulacastrum in Escherichia coli

本研究利用从海马齿中克隆到的甜菜碱醛脱氢酶基因构建到高效表达载体p GEX-4T-1上,构建重组载体p GEX-4T-Sp BADH,并对GST-Sp BADH融合表达的IPTG诱导浓度、诱导不同温度、不同菌液浓度和诱导时间等条件进行优化。研究结果表明:随诱导时间增长GST-Sp BADH融合蛋白表达量提高,在37℃时,OD为0.6左右,诱导5 h GST-Sp BADH融合蛋白的表达量达到最大,在0.2 mmol/L IPTG浓度下,可以有效诱导GST-Sp BADH融合蛋白的表达。本研究结果以期解析Sp BADH基因的功能以及在海马齿抗盐过程中的作用机理。

A full-length betaine aldehyde dehydrogenase gene sequence ofSesuvium portulaeastrum was cloned into the high expression vector pGEX-4T-1, and named pGEX-4T-SpBA DH. The GST-SpBADH fusion protein was expressed induced by IPTG. The expression conditions such as IPTG concentration, the bacterium concen- tration, induction time and temperature were optimized. The results showed that the expression level of GST- SpBADH was increased by accompanying with the induction time. In the condition of 37℃, OD600 is 0.6 and cultured 5 hours, GST-SpBADH fusion protein reached the highest expression. 0.2 mmol/L concentration of IPTG can induce GST-SpBADH expression effectively in Eseherichia coli expression system. The aim of this research was to analyze the function and salt tolerance mechanism of SpBA DH gene.