中国明对虾ALFFc基因的原核表达与抗血清制备

利用聚合酶链式反应(PCR)技术和基因重组技术克隆了中国明对虾(Fenneropenaeus chinensis)抗脂多糖因子(ALFFc)基因的成熟肽编码序列并构建了pET-DsbA-ALFFc原核表达载体,转化E.coli BL21(DE3)后,经异丙基-β-D-硫代半乳糖苷(IPTG)筛选法获得了1株高表达量重组菌,并通过实验确定其最佳诱导条件为:菌液初始OD6000.7,IPTG终浓度0.8 mmol/L,培养温度37℃,诱导时间3 h.重组蛋白经纯化后免疫新西兰大白兔制备了抗血清,为进一步研究ALFFc的功能奠定了基础.

Prokaryotic Expression and Antiserum Preparation of ALFFc in Chinese Shrimp (Fenneropenaeus chinensis)

The sequence coding mature peptide of anti-lipopolysaccharide factor(ALFFc)was cloned from Chinese shrimp Fenneropenaeus chinensis,and a prokaryotic expression vector pET-DsbA-ALFFc was constructed by PCR and recombinant DNA techniques.A high-productivity colony of transformed E.coli BL21(DE3) was obtained by IPTG screening method.The optimal conditions for the introduction of the fused protein were investigated,which are described as follows: final IPTG concentration is 0.8 mmol/L,initial OD600 is 0.7,and ...