| 中国明对虾ALFFc基因的原核表达与抗血清制备 |
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| 引用本文： | 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) |
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| 作者姓名： | 柳峰松 李婷 王晓菲 唐婷 |
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| 作者单位： | 河北大学,生命科学学院,河北,保定,071002;河北大学,生命科学学院,河北,保定,071002;河北大学,生命科学学院,河北,保定,071002;河北大学,生命科学学院,河北,保定,071002 |
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| 基金项目： | 河北省自然科学基金资助项目(C2008000596) |
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| 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.
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| 关 键 词： | 中国明对虾 抗脂多糖因子(ALFFc) 原核表达 抗血清 |
Prokaryotic Expression and Antiserum Preparation of ALFFc in Chinese Shrimp (Fenneropenaeus chinensis) |
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| 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) |
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| Authors: | LIU Feng-song LI Ting WANG Xiao-fei TANG Ting |
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| Affiliation: | College of Life Sciences;Hebei University;Baoding 071002;China |
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| 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 OD600 is 0.7,and ... |
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| Keywords: | Fenneropenaeus chinensis ALFFc prokaryotic expression antiserum |
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