【摘要】 为研究乙烯信号转录因子的功能,提取破色期番茄果实总RNA,通过RT-PCR获得615bp全长LeERF1基因。测序结果表明,与已发表序列同源性为100%。将LeERF1基因亚克隆至载体pET-30a上,构建原核表达载体pET-LeERF1并转化到大肠杆菌BL21中。采用诱导温度37℃,诱导剂IPTG浓度1mmol/L,诱导时间3h,LeERF1表达蛋白在细菌中占菌体总蛋白的28%。更多还原

【Abstract】 In order to study the function of ethylene signal transcription factor,the 615 bp entire coding sequence of LeERF1 was cloned by RT-PCR from breaking tomato fruit RNA. Sequential determination result showed that the identity was 100% between cloned gene and published gene. The PCR product was subcloned into prokaryotic protein expression vector pET-30a to generate recombinant plasmid pET-LeERF1 in E. coli BL 21. LeERF1 protein was expressed under 1.0 mmol/L IPTG induction for 3 h at 37 ℃. The expression level of LeERF1 was about 28% of total cellular proteins.更多还原