中国明对虾ALFFc基因的原核表达与抗血清制备  

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

作　　者：柳峰松[1] 李婷[1] 王晓菲[1] 唐婷[1] 

机构地区：[1]河北大学生命科学学院,河北保定071002

出　　处：《河北大学学报（自然科学版）》2010年第1期88-92,共5页Journal of Hebei University(Natural Science Edition)

基　　金：河北省自然科学基金资助项目(C2008000596)

摘　　要：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.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 introduction is conducted at 37 ℃ for 3 h. The purified recombinant protein DsbA-ALFFc was used as antigen to prepare antiserum in New Zealand white rabbit, which provides the basis for further functional analysis of ALFFc.

关 键 词：中国明对虾 抗脂多糖因子(ALFFc) 原核表达 抗血清 

分 类 号：Q78[生物学—分子生物学]