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题名	抗玉米素核苷的单克隆抗体的制备及植物内源细胞分裂素的酶联免疫吸附测定
作者	张龙
学位类别	硕士
答辩日期	1988
授予单位	中国科学院昆明植物研究所
授予地点	中国科学院昆明植物研究所
导师	季本仁
中文摘要	通过二次成功的细胞融合，共获得三株分泌抗玉米素核苷（ZR）的单克隆抗体的稳定杂交瘤细胞系（F_2E_6、F_3G_6、F_1B_9），其中F_2E_6分泌的单抗属IgG_1，F_5G_6，F_1B_9，分泌的单抗均属IgM，以F_2E_6，F_3G_6单抗为基础的间接酶联免疫吸附试法（ELISA）对ZR的检测灵敏度为0.05 Pmol（～18Pg），线性检测区界为0.05-50 Pmol。反式玉米素（t-z），顺式玉米素（c-z），异戊烯基腺苷（IPA）、6-苄基腺嘌呤（6-BA）、激动素（KT）和腺苷与ZR的竟争反应曲线表明，除玉米素以外，其它几种细胞分裂素的交叉反应值都低于0.18%，因此，F_3G_6、F_2E_6单抗具高度的特异性，用ELISA测定F_2E_6单抗与玉米素核苷反应的亲合常数为2.36 ± 0.99 * 10~(-8)M，故此单抗与ZR有较强的亲合力，本文还对以单抗为基础的包埋抗原的ELISA在植物内源细胞分裂素测定中的应用进行了研究，并用此方法测定了黄化玉米（Zea、mays L.）幼苗中玉米素核苷类的含量。结果表明，ELISA法不但具有高度的选择和灵敏度，样品提取液只需简单处理即可进行测定，无任何干扰物质存在。而且具有操作简便，安全、周期短（约3小时），不需任何昂贵仪器，同时可处理大量样品等优点，因此是一种值得推广的激素测定方法。
英文摘要	Three hybridomas, F_2E_6, F_1B_9, F_3G_6, which secreted monoclonal anti-zeatin riboside antibodies, were produced from two successive fusions of BALB/c spleen cells with Sp2/0 myeloma cells. The mice had been hyperimmunized with zeatin riboside-bovine serum albumin conjugate for about 6 months and the positive hybridomas were screened by indirect enzyme-linked immuno-absorbent assay. Among these hybridomas, F_2E_6 secretes antibody of the IgG_1, both F_3G_6, F_1B_9 secrete the IgM. Two of the monoclonal antibodies, F_2E_6, F_3G_6, were characterized by ELISA and found that the detection limits were 0.05pmol(～18pg) of zeatin riboside and the measuring ranges extended from 0.05-50pmol. Competitive binding trials against trans-Zeatin, cis-Zeatin, Isopentenyladenosine, 6-Benzylaminopurine, 6-Furfurylaminopurine, and Adenosine showed that except Zeatin, the cross-reactivities towards these structurally related cytokinins were negligible (less than 0.18%). So these monoclonal antibodies have high specificity. The affinity constant of F_2E_6 monoclonal antibody, measured by ELISA, was 2.36 * 10~(-8)M, which indicates that this monoclonal antibody has high affinity for zeatin riboside. The use of immobilized antigen ELISA in plant cytokinin detection were discussed and the content of cytokinins (calculated as zeatin riboside) in etiolated Zea mays L. seedling extracts was analysed with a little handlement. The method reported here not only has high sensitivity and specificity, but also has the advantages of being simpler, safer, time-saving (3 hours), requiring less expensive equipment and handling a lots of samples at the same time, so be worth spreading vastly.
语种	中文
公开日期	2011-10-25
页码	57
内容类型	学位论文
源URL	[http://ir.kib.ac.cn/handle/151853/820]
专题	昆明植物研究所_昆明植物所硕博研究生毕业学位论文
推荐引用方式 GB/T 7714	张龙. 抗玉米素核苷的单克隆抗体的制备及植物内源细胞分裂素的酶联免疫吸附测定[D]. 中国科学院昆明植物研究所. 中国科学院昆明植物研究所. 1988.

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