| ING4的原核表达及多克隆抗体的制备 |
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| 引用本文： | 王金志,缪竞诚,盛伟华,谢宇锋,杨吉成.ING4的原核表达及多克隆抗体的制备[J].苏州大学学报(自然科学版),2006,26(6):930-932,935. |
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| 作者姓名： | 王金志 缪竞诚 盛伟华 谢宇锋 杨吉成 |
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| 作者单位： | 苏州大学医学院基础医学系细胞与分子生物学教研室,江苏苏州215123 |
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| 摘 要： | 目的在原核系统中表达并纯化ING4蛋白，制备多克隆抗体。方法构建表达载体pET-21a（＋）-ING4，转化大肠杆菌BL21（DE3），用IPTG诱导表达。SDS-PAGE电泳检测。结果构建的表达载体pET-21a（＋）-hGDNF经酶切、PCR鉴定均出现750bp左右的条带，测序结果正确，用IPTG诱导转化菌有30KD大小的目的条带，表达的目的蛋白占菌体总蛋白10％以上，以包涵体形式存在，纯化后其纯度可达70％以上。用纯化的ING4蛋白免疫新西兰兔制备多克隆抗体。结论制备的多克隆抗体为下阶段的研究提供了重要的实验材料。
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| 关 键 词： | 原核表达 抗体 |
| 文章编号： | 1673-0399（2006）06-0930-03 |
| 收稿时间： | 2005-11-05 |
| 修稿时间： | 2005-11-05 |
Prokaryotic Expression of ING4 and the Preparation of Polyclonal Antibody |
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| WANG JIN-zhi, MIAO JING-cheng, SI-LENG WEI-hua, et al.Prokaryotic Expression of ING4 and the Preparation of Polyclonal Antibody[J].Suzhou University Journal of Medical Science,2006,26(6):930-932,935. |
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| Authors: | WANG JIN-zhi MIAO JING-cheng SI-LENG WEI-hua |
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| Institution: | Depf of Celluar and Molecular Biology, Medical College of Suzhou University, Jiangsu Suzhou 215123, China |
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| Abstract: | Objective To express and purify the recombinant ING4in prokaryotic system and to prepare polyclonal antibody.Methods The E.coli expression vector pET-21a(+)-ING4was constructed and transferred into E.coli BL21(DE3).The expression of ING4 protein in transformant was controled by T7 promotor with IPTG,the products was detected by SDS-PAGE.Results Recombiant protein was expressed in E.coli host strain BL21(DE3) in the form of inclusion body,and the expression level was 10% higher than that in the total cell lysate.The ING4 amounted to at least 80% after purification.Conclusion The polyclonal antibody can be used in further studies. |
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| Keywords: | ING4 |
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