中国明对虾ALFFc基因的原核表达与抗血清制备

doi:10.3969/j.issn.1000-1565.2010.01.019

摘要/Abstract

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OD_(600) 0.7, 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.

关键词: 中国明对虾, 抗脂多糖因子(ALFFc), 原核表达, 抗血清

Abstract: 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 OD_(600) 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.

Key words: Fenneropenaeus chinensis, ALFFc, prokaryotic expression, antiserum

中图分类号:

柳峰松,李婷,王晓菲,唐婷. 中国明对虾ALFFc基因的原核表达与抗血清制备[J]. 河北大学学报(自然科学版), 2010, 30(1): 88-92.

LIU Feng-song,LI Ting,WANG Xiao-fei,TANG Ting. Prokaryotic Expression and Antiserum Preparation of ALFFc in Chinese Shrimp (Fenneropenaeus chinensis)[J]. Journal of Hebei University (Natural Science Edition), 2010, 30(1): 88-92.

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