Shimizu N, Shimizu Y, Kucherlapati R S, Ruddle F H
Cell. 1976 Jan;7(1):123-30. doi: 10.1016/0092-8674(76)90262-2.
Rabbit antisera have been produced against each of three purified human enzymes: a cytoplasmic form of NADP-linked isocitrate dehydrogenase (IDH, EC 1.1.1.42), phosphoglucose isomerase (PGI, EC 5.3.1.9), and hypoxanthine guanine phosphoribosyltransferase (HGPRT, EC 2.4.2.8), and they have been used for immunoprecipitation reactions to detect human-specific enzymes in various human-mouse somatic cell hybrids. Under optimal conditions, enzyme activity was eliminated from human cell lysate, but no reduction of enzyme activity was found in the mouse cell lysate. Differential enzyme precipitation by these human-specific antisera was observed in human-mouse hybrid cells. Analysis on starch gel electrophoresis revealed that not only the human homodimer, but also human-mouse heterodimer molecules, in cases of PGI and IDH, were precipitated. Thus this method is sensitive and allows quantitative determination of human-specific enzymes. The presence of a human-specific enzyme identified by this method correlated with the presence of a particular human chromosome permitting assignments of the human cytoplasmic forms of NADP-linked IDH, human PGI, and human HGPRT genes to chromosomes 2, 19, and X, respectively. These assignments are consistent with published data (Ruddle, 1973).
一种胞质形式的NADP连接的异柠檬酸脱氢酶（IDH，EC 1.1.1.42）、磷酸葡萄糖异构酶（PGI，EC 5.3.1.9）和次黄嘌呤鸟嘌呤磷酸核糖转移酶（HGPRT，EC 2.4.2.8），并将其用于免疫沉淀反应，以检测各种人-小鼠体细胞杂种中的人类特异性酶。在最佳条件下，人细胞裂解物中的酶活性被消除，但在小鼠细胞裂解物中未发现酶活性降低。在人-小鼠杂种细胞中观察到这些人类特异性抗血清对酶的差异性沉淀。淀粉凝胶电泳分析表明，对于PGI和IDH，不仅人同源二聚体，而且人-小鼠异源二聚体分子都被沉淀。因此，该方法灵敏，可对人类特异性酶进行定量测定。通过该方法鉴定出的人类特异性酶的存在与特定人类染色体的存在相关，从而分别将NADP连接的IDH、人类PGI和人类HGPRT基因的人类胞质形式定位到2号、19号和X染色体上。这些定位与已发表的数据一致（Ruddle，1973年）。
