Miller C L, Ruddle F H
Proc Natl Acad Sci U S A. 1978 Jul;75(7):3346-50. doi: 10.1073/pnas.75.7.3346.
Transformation frequencies of 4 x 10(-5) were obtained in chromosome-mediated gene transfer experiments using human cell line HeLa S3 as donor and mouse cell line A9 as recipient. This high frequency of interspecific transformation was achieved by treating the recipient cells with dimethylsulfoxide in addition to other facilitators. The high frequency of transformation correlated positively with transgenome size on the basis of both co-transfer of linked markers and chromosome analysis. The syntenic human markers glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate:NADP(+) 1-oxidoreductase, EC 1.1.1.49) and phosphoglycerate kinase (ATP:3-phospho-D-glycerate 1-phosphotransferase, EC 2.7.2.3) were sometimes transferred together with the selected X-linked prototrophic marker hypoxanthine phosphoribosyltransferase (IMP: pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) into murine somatic cells. Donor human chromosome material could be demonstrated cytologically in some of the transformed cell lines. Transformants exhibited various rates of loss of the human hypoxanthine phosphoribosyltransferase marker when grown under nonselective conditions. These results reveal a broader range of possible interspecific transgenome sizes than has been recognized in the past. The largest transgenomes consist of cytologically detectable donor fragments and contain syntenic markers that are not closely linked to the selected marker.
在以人类细胞系HeLa S3作为供体、小鼠细胞系A9作为受体的染色体介导的基因转移实验中，获得了4×10⁻⁵的转化频率。除了其他促进剂外，通过用二甲基亚砜处理受体细胞，实现了这种高频率的种间转化。基于连锁标记的共转移和染色体分析，转化的高频率与转基因组大小呈正相关。同线的人类标记葡萄糖-6-磷酸脱氢酶（D-葡萄糖-6-磷酸：NADP⁺ 1-氧化还原酶，EC 1.1.1.49）和磷酸甘油酸激酶（ATP：3-磷酸-D-甘油酸1-磷酸转移酶，EC 2.7.2.3）有时会与选定的X连锁原养型标记次黄嘌呤磷酸核糖基转移酶（IMP：焦磷酸磷酸核糖基转移酶，EC 2.4.2.8）一起转移到小鼠体细胞中。在一些转化细胞系中，从细胞学上可以证明存在供体人类染色体物质。当在非选择性条件下生长时，转化体表现出人类次黄嘌呤磷酸核糖基转移酶标记的不同丢失率。这些结果揭示了比过去所认识到的更广泛的可能种间转基因组大小范围。最大的转基因组由细胞学上可检测到的供体片段组成，并包含与选定标记没有紧密连锁的同线标记。
