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识别禽肉瘤病毒转化蛋白的单克隆抗体的分离

Isolation of monoclonal antibodies that recognize the transforming proteins of avian sarcoma viruses.

作者信息

Lipsich L A, Lewis A J, Brugge J S

出版信息

J Virol. 1983 Nov;48(2):352-60. doi: 10.1128/JVI.48.2.352-360.1983.

DOI:10.1128/JVI.48.2.352-360.1983
PMID:6312092
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC255359/
Abstract

Thirteen clones of hybrid cells which synthesize antibodies directed against the Rous sarcoma virus (RSV) transforming protein, pp60src, were isolated. Mouse myeloma cells were fused with spleen cells from mice that had been immunized with purified pp60src from bacterial recombinants which direct the synthesis of the RSV src gene. The hybridomas which survived the selection medium were screened by immunoprecipitation of pp60src from 32P-labeled lysates of RSV-transformed cells. Monoclonal antibodies produced by subclones derived from 13 hybridomas recognized pp60src encoded by the Schmidt-Ruppin and Prague strains of RSV and the cellular homolog of pp60src. Antibody from clone 261 had a high affinity for the viral yes gene product, and antibodies from clones 443 and 463 recognized the transforming proteins encoded by viruses containing the related transforming genes fps and ros. Several other clones had a low affinity for the viral yes, fps, and ros gene products which could be detected by in vitro phosphorylation of the transforming proteins after immunoprecipitation with the monoclonal antibody. All of the monoclonal antibodies allowed phosphorylation of pp60src and casein in an immune complex-bound reaction.

摘要

分离出了13个合成针对劳氏肉瘤病毒（RSV）转化蛋白pp60src的抗体的杂交细胞克隆。将小鼠骨髓瘤细胞与来自用细菌重组体中指导RSV src基因合成的纯化pp60src免疫的小鼠的脾细胞融合。在选择培养基中存活下来的杂交瘤通过从RSV转化细胞的32P标记裂解物中免疫沉淀pp60src进行筛选。来自13个杂交瘤的亚克隆产生的单克隆抗体识别由RSV的施密特-鲁平株和布拉格株编码的pp60src以及pp60src的细胞同源物。来自克隆261的抗体对病毒yes基因产物具有高亲和力，来自克隆443和463的抗体识别由含有相关转化基因fps和ros的病毒编码的转化蛋白。其他几个克隆对病毒yes、fps和ros基因产物具有低亲和力，在用单克隆抗体免疫沉淀后，通过转化蛋白的体外磷酸化可以检测到这种低亲和力。所有单克隆抗体在免疫复合物结合反应中都能使pp60src和酪蛋白磷酸化。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/572a1ce665d2/jvirol00140-0029-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/ca76d00066e5/jvirol00140-0026-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/dc0c3e11e6e9/jvirol00140-0027-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/90d8b3bb1463/jvirol00140-0028-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/eb12962a2c37/jvirol00140-0028-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/572a1ce665d2/jvirol00140-0029-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/ca76d00066e5/jvirol00140-0026-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/dc0c3e11e6e9/jvirol00140-0027-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/90d8b3bb1463/jvirol00140-0028-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/eb12962a2c37/jvirol00140-0028-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7dc8/255359/572a1ce665d2/jvirol00140-0029-a.jpg

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Identification and characterization of cellular targets for tyrosine protein kinases.酪氨酸蛋白激酶细胞靶点的鉴定与表征
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Development of anti-pp60src serum with antigen produced in Escherichia coli.利用在大肠杆菌中产生的抗原制备抗pp60src血清。
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