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通过分子克隆和DNA介导的基因转移实现β2-微球蛋白与ly-m11的连锁。

Linkage of beta 2-microglobulin and ly-m11 by molecular cloning and DNA-mediated gene transfer.

作者信息

Margulies D H, Parnes J R, Johnson N A, Seidman J G

出版信息

Proc Natl Acad Sci U S A. 1983 Apr;80(8):2328-31. doi: 10.1073/pnas.80.8.2328.

Abstract

beta 2-Microglobulin (beta 2m) is expressed on the cell surface after introduction of a beta 2mb (C57BL/6N) genomic clone into thymidine kinase-deficient mouse L cells by cotransformation using the calcium phosphate precipitate method. Stable transformant cell lines were identified that express the beta 2mb allele, as determined by reaction of the cells with appropriate monoclonal antibodies and by two-dimensional gel electrophoresis of endogenously labeled immunoprecipitates of cell extracts. These beta 2mb transformants now express ly-m11.2, as detected by an indirect radioimmunoassay. A plasmid subclone of the beta 2mb gene that contains an 8.4-kilobase insert, after introduction into mouse L cells, similarly directs the synthesis of both the beta 2mb and the ly-m11.2 antigens. Thus, the beta 2mb and ly-m11.2 determinants most likely represent sites on the same protein structure.

摘要

通过使用磷酸钙沉淀法共转化,将β2m(C57BL/6N)基因组克隆导入胸苷激酶缺陷型小鼠L细胞后,β2-微球蛋白(β2m)在细胞表面表达。通过细胞与适当的单克隆抗体反应以及对细胞提取物中内源性标记免疫沉淀物的二维凝胶电泳分析,鉴定出表达β2m等位基因的稳定转化细胞系。通过间接放射免疫测定法检测到,这些β2m转化体现在表达ly-m11.2。将包含8.4千碱基插入片段的β2m基因质粒亚克隆导入小鼠L细胞后,同样指导β2m和ly-m11.2抗原的合成。因此,β2m和ly-m11.2决定簇很可能代表同一蛋白质结构上的位点。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b15a/393813/3413b04180f1/pnas00634-0240-a.jpg

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