人类Foxm1b基因的原核表达、抗体制备及其对肝癌细胞中Foxm1b蛋白的识别
【目的】构建人类Foxm1b基因的原核表达载体,表达并纯化蛋白,制备多克隆抗体,并用此抗体检测肝癌细胞中Foxm1b基因的表达,为进一步研究Foxm1b基因的功能奠定基础。【方法】从GenBank中下载人类Foxm1b基因全长cDNA序列并用Pegene软件分析其可能的抗原表位,设计引物.应用RT—PCR获得含抗原表位的Foxm1b基因片段,重组人原核表达载体pET-28a,在大肠杆菌BL-21(DE31中诱导表达,应用亲和层析法获得纯度较高的原核表达蛋白并免疫大鼠制备多克隆抗体,用ELISA检测抗体滴度并用免疫细胞化学的方法检测此抗体对肝癌细胞中天然Foxm1b蛋白的识别。【结果】成功构建了...
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Main Authors: | 唐保东, 刘思纯, 徐雅, 曾志荣, 胡品津 |
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Format: | Article |
Language: | zho |
Published: |
Editorial Office of Journal of Sun Yat-sen University
2007-01-01
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Series: | Zhongshan Daxue xuebao. Yixue kexue ban |
Subjects: | |
Online Access: | http://xuebaoyx.sysu.edu.cn/zh/article/43581731/ |
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