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针对兔痘病毒相关的DNA指导的RNA聚合酶两个亚基的单克隆抗体的分离与鉴定。

Isolation and characterization of monoclonal antibodies directed against two subunits of rabbit poxvirus-associated, DNA-directed RNA polymerase.

作者信息

Morrison D K, Carter J K, Moyer R W

出版信息

J Virol. 1985 Sep;55(3):670-80. doi: 10.1128/JVI.55.3.670-680.1985.

DOI:10.1128/JVI.55.3.670-680.1985
PMID:2991589
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC255039/
Abstract

A library of monoclonal antibodies directed against individual proteins of the rabbit poxvirus (RPV) virion within a complex immunogenic mixture has been generated through the use of in vivo and in vitro immunization regimens. The relative efficacies of the two procedures were compared. Based on immunoblot analysis, the in vitro immunization regimen led both to a wider variety of monoclonal antibodies to different proteins and to a larger number of antibodies directed against proteins of higher molecular weights. Each method, however, has advantages, and the two procedures appear to be complementary. A simple method to recognize antibodies directed against the virion DNA-directed RNA polymerase was developed. Monoclonal antibodies directed against two subunits (137 and 34 kilodaltons [kDa]) of the RNA polymerase were identified and used to study the biogenesis of the enzyme and to map the two corresponding genes within the viral genome by using an RPV DNA library cloned into the lambda gtll expression vector. Both proteins are synthesized late in the infectious cycle and are restricted totally to the cytoplasm. Preliminary mapping data place the genes encoding the 137-kDa protein within the HindIII H fragment, whereas the gene for the 34-kDa protein is located within the left most region of the HindIII A fragment.

摘要

通过体内和体外免疫方案，已构建了一个针对兔痘病毒（RPV）病毒粒子中单个蛋白质的单克隆抗体文库，该文库存在于复杂的免疫原性混合物中。比较了这两种方法的相对有效性。基于免疫印迹分析，体外免疫方案产生了针对不同蛋白质的更多种类的单克隆抗体，以及更多针对高分子量蛋白质的抗体。然而，每种方法都有其优点，这两种方法似乎具有互补性。开发了一种识别针对病毒粒子DNA指导的RNA聚合酶的抗体的简单方法。鉴定了针对RNA聚合酶两个亚基（137和34千道尔顿[kDa]）的单克隆抗体，并用于研究该酶的生物发生，并通过使用克隆到λgtll表达载体中的RPV DNA文库在病毒基因组中定位两个相应的基因。这两种蛋白质均在感染周期后期合成，并且完全局限于细胞质中。初步定位数据表明，编码137-kDa蛋白质的基因位于HindIII H片段内，而34-kDa蛋白质的基因位于HindIII A片段最左侧区域。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/6564852cd747/jvirol00120-0172-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/57ad457c77e1/jvirol00120-0167-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/67f6af2c8152/jvirol00120-0169-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/ac3bfe17a32b/jvirol00120-0170-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/fb0877059295/jvirol00120-0171-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/12db153efe39/jvirol00120-0171-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/9bfbf8a2b82b/jvirol00120-0171-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/6564852cd747/jvirol00120-0172-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/57ad457c77e1/jvirol00120-0167-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/67f6af2c8152/jvirol00120-0169-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/ac3bfe17a32b/jvirol00120-0170-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/fb0877059295/jvirol00120-0171-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/12db153efe39/jvirol00120-0171-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/9bfbf8a2b82b/jvirol00120-0171-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/52cd/255039/6564852cd747/jvirol00120-0172-a.jpg

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