高等学校化学学报

高等学校化学学报 ›› 2001, Vol. 22 ›› Issue (3): 403.

• 研究论文 • 上一篇    下一篇

低分子量紫色酸性磷酸酶多克隆抗体的制备及应用

范洪宽1, 郑远志2, Hamilton Susan2, de Jersery John2   

  1. 1. 吉林大学生命科学学院, 长春 130023;
    2. DepartmentofBiochemistry, TheUniversityofQueensland, Brisbane, Australia 4072
  • 收稿日期:2000-04-06 出版日期:2001-03-24 发布日期:2001-03-24
  • 通讯作者: 范洪宽(1973年出生),男,28岁,博士研究生,从事FGF拮抗剂研究.E-mail:liwei@mai.ljlu.edu.cn E-mail:liwei@mai.ljlu.edu.cn

Generation and Application of Polyclonal Antibodies Against Low Molecular Weight Purple Acid Phosphatase

FAN Hong-Kuan1, ZHENG Yuan-Zhi2, Hamilton Susan2, de Jersey John2   

  1. 1. College of Life Science, Jilin University, Changchun 130023, China;
    2. Department of Biochemistry, The University of Queensland, Brisbane 4072, Australia
  • Received:2000-04-06 Online:2001-03-24 Published:2001-03-24

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被引次数

8

摘要： 将最近从地瓜中提取出来的低分子量紫色酸性磷酸酶(smPAP)基因克隆到GST融合蛋白表达载体pGEX-2T中,在大肠杆菌 BL21 codonplus中进行表达,用表达的融合蛋白免疫兔子产生多克隆抗体.用抗血清对昆虫细胞中表达的smPAP和地瓜提取液中分离纯化的PAP进行检测,产生了良好的交叉反应.

关键词: 紫色酸性磷酸酶, 融合蛋白, 多克隆抗体

Abstract: There exists two groups of purple acid phosphatases(PAPs). Mammalian PAPs, which are monomers with molecular weight in the range of 33000-40000. Plant PAPs, which are dimers or tetramers containing identical subunits of about 55000. Recently the gene of low molecular weight PAPs(smPAPs) was found in plant. They are significantly similar to mammalian PAPs' but quite different from plant PAPs'. In order to understand the exact function of the protein and its distribution in plant, we cloned the sweet potato smPAPgene in pGEX-2T vector and expressed the fusion protein in E. coli. The expressed fusion protein was used to generate polyclonal antibodies. The antibodies demonstrated a good reactivity towards the smPAPexpressed in baculovirus and PAPs partially purified from sweet potato juice. Therefore, the antibodies make it possible to purify and characterize the smPAP.

Key words: Purple acid phosphatase, Fusion protein, Polyclonal antibodies

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