Fell H P, Yarnold S, Hellström I, Hellström K E, Folger K R
Oncogen, Seattle, WA 98121.
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8507-11. doi: 10.1073/pnas.86.21.8507.
We demonstrate that murine myeloma cells can efficiently mediate homologous recombination. The murine myeloma cell line J558L was shown to appropriately recombine two transfected DNA molecules in approximately 30% of cells that received and integrated intact copies of both molecules. This activity was then exploited to direct major reconstructions of an endogenous locus within a hybridoma cell line. Production of antigen-specific chimeric heavy chain was achieved by targeting the human IgG1 heavy chain constant region (C gamma 1) exons to the genomic heavy chain locus of a hybridoma cell line secreting antibody specific for a human tumor-associated antigen. The frequency of productive genomic recombinations was approximately 1 in 200 transfectants, with accumulation of the chimeric protein reaching greater than 20 micrograms/ml in culture supernatants.
我们证明小鼠骨髓瘤细胞能够有效地介导同源重组。小鼠骨髓瘤细胞系J558L在约30%接收并整合了两个分子完整拷贝的细胞中，显示出能恰当地重组两个转染的DNA分子。然后利用这种活性来指导杂交瘤细胞系内一个内源性位点的重大改造。通过将人IgG1重链恒定区（Cγ1）外显子靶向到一个分泌针对人肿瘤相关抗原的特异性抗体的杂交瘤细胞系的基因组重链位点，实现了抗原特异性嵌合重链的产生。有生产性的基因组重组频率约为每200个转染子中有1个，培养上清液中嵌合蛋白的积累量达到大于20微克/毫升。
