The use of cell surface antigens to characterize and select for fragments of human chromosomes retained by interspecies hybrids.
The use of cell surface antigens to characterize and select for fragments of human chromosomes retained by interspecies hybrids.
复制标题
使用细胞表面抗原来表征和选择种间杂交保留的人类染色体片段。
DOI：
10.1002/jez.1402300211
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发表时间：
1984
期刊：
影响因子：
--
通讯作者：
Ruddle,FH
中科院分区：
文献类型：
--
作者：
Kamarck,ME;Macyko,CA;Jones,C;Ruddle,FH
我们使用了通过人类染色体介导的基因转移（CMGT）产生的小鼠细胞悬液，以探索细胞表面抗原在鉴定体细胞杂交体保留的人类染色体片段中的用途。转化株系21- 30 b含有完整的重排的人染色体，同工酶分析表明含有来自9号和X号染色体的遗传物质。通过在小鼠中使用该抗体作为免疫原，还可以产生针对人类特异性表面抗原的抗血清。利用人×啮齿类动物杂交细胞系，控制这些抗原表达的基因可定位于11 pter → 11 p13，与表面抗原S3一致分离。尽管染色体特异性同工酶LDH-A或表面抗原W 6/34和4F 2的存在无法检测到染色体11在大肠杆菌中的存在，但这些结论是可能的。最后，使用荧光激活细胞分选仪（FACS）将PBMC细胞破碎成抗原阳性和阴性亚群。这导致了四个额外的染色体重排涉及种间染色体易位的分离和表征。这项工作证明了染色体特异性表面抗原和FACS在评价种间杂种保留的人类染色体片段中的价值。
We have used a mouse cell transformant generated by human chromosome‐mediated gene transfer (CMGT) to explore the use of cell surface antigens in the identification of fragments of human chromosomes retained by somatic cell hybrids. The transformed line, 21–30b, contained an intact rearranged human chromosome, and could be shown by isozyme analysis to contain genetic material from chromosomes 9 and X. By using the transformant as an immunogen in mice, it was also possible to produce antiserum to human‐specific surface antigens. Using genetically characterized human × rodent hybrid lines, the genes controlling expression of these antigens could be localized to 11pter → 11p13, segregating concordantly with surface antigen S3. These conclusions were possible despite the fact that the presence of chromosome 11 in the transformant was not detectable by the presence of chromosome specific isozyme LDH‐A or surface antigens W6/34 and 4F2. Finally, the fluorescence‐activated cell sorter (FACS) was used to fractionate the transformant cells into antigen positive and negative subpopulations. This resulted in the isolation and characterization of four additional chromosome rearrangements involving interspecies chromosome translocations. This work demonstrates the value of chromosome‐specific surface antigens and the FACS in the evaluation of human chromosome fragments retained by interspecies hybrids.