| 凡纳滨对虾铁蛋白基因的克隆与表达 |
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| 引用本文： | 吴文林,傅玲玲,陈宇,陈曦.凡纳滨对虾铁蛋白基因的克隆与表达[J].泉州师范学院学报,2011,29(6):7-10. |
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| 作者姓名： | 吴文林 傅玲玲 陈宇 陈曦 |
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| 作者单位： | 泉州师范学院化学与生命科学学院,福建泉州,362000 |
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| 基金项目： | 国家自然科学基金(31072236); 福建省教育厅项目(JK2009043) |
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| 摘 要： | 以凡纳滨对虾为研究对象，通过RT-PCR技术扩增出铁蛋白（Ferritin）基因，经特定的酶切（Notl，Smal）后插入到表达质粒pGEX-4T-2上构建表达载体，重组质粒转化大肠杆菌．经菌落PCR和质粒双酶切鉴定确认，证实成功地构建了对虾铁蛋白基因的原核表达载体．再对重组菌用IPTG诱导表达，采用Glutathione Sepharose4B亲和层析得到目的蛋白用于制备抗体，为研究铁蛋白在对虾抗病毒免疫中的作用奠定基础．
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| 关 键 词： | 凡纳滨对虾 铁蛋白 重组 表达 |
Cloning,Expression and Purification of Ferritin from Litopenaeus Vannamei |
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| WU Wen-lin,FU Ling-ling,CHEN Yu,CHEN Xi.Cloning,Expression and Purification of Ferritin from Litopenaeus Vannamei[J].Journal of Quanzhou Normal College,2011,29(6):7-10. |
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| Authors: | WU Wen-lin FU Ling-ling CHEN Yu CHEN Xi |
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| Institution: | WU Wen-lin,FU Ling-ling,CHEN Yu,CHEN Xi(School of Chemistry and Life Science,Quanzhou Normal University,Fujian 362000,China) |
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| Abstract: | In this study, ferritin gene from Litopenaeus vannamei was amplified by PCR, digested by Sma I and Not I, and inserted into the expression vector pGEX-4T-2 to construct recombinant plasmid. Colonies with recombinant plasmid were picked for PCR and culture identified by enzyme digestion. The results show that ferritin gene ean be successfully inserted into the pGEX-4T-2 expression vector. After induced with IPTG at 18℃, the fusion protein can be purified with Glutathione Sepharose 4B. |
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| Keywords: | Litopenaeus vannamei ferritin recombination expression |
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