Margulies D H, Ramsey A L, Boyd L F, McCluskey J
Proc Natl Acad Sci U S A. 1986 Jul;83(14):5252-6. doi: 10.1073/pnas.83.14.5252.
We have constructed a recombinant class I gene in which 5' sequences of H-2Dd are linked to the 3' half of a Qa subregion gene, Q10b. This hybrid gene would be expected to direct the synthesis of a protein containing the N and C1 domains of H-2Dd covalently linked to the C2 domain of the secreted, nonpolymorphic, Q10b antigen. Following DNA-mediated gene transfer into mouse L cells, transformants were analyzed by radiolabeling and immunoprecipitation. These cells secreted a molecule reactive with anti-H-2Dd monoclonal antibodies that identify epitopes on the N and C1 domains as well as with an anti-Q10 carboxyl-terminal peptide antiserum. The H-2Dd-derived antigen is associated with beta 2-microglobulin and is readily purified in milligram amounts from culture supernatants by immunoaffinity chromatography.
我们构建了一个重组的I类基因，其中H-2Dd的5'序列与Qa亚区基因Q10b的3'半段相连。预计这个杂合基因会指导合成一种蛋白质，该蛋白质包含H-2Dd的N和C1结构域，并与分泌型、非多态性的Q10b抗原的C2结构域共价连接。在将DNA介导的基因转移到小鼠L细胞后，通过放射性标记和免疫沉淀对转化体进行了分析。这些细胞分泌出一种能与抗H-2Dd单克隆抗体发生反应的分子，该抗体可识别N和C1结构域上的表位，同时也能与抗Q10羧基末端肽抗血清发生反应。源自H-2Dd的抗原与β2-微球蛋白相关，并且可以通过免疫亲和色谱法从培养上清液中轻松纯化出毫克量的该抗原。
