Huxley C, Hagino Y, Schlessinger D, Olson M V
Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110.
Genomics. 1991 Apr;9(4):742-50. doi: 10.1016/0888-7543(91)90369-p.
A 680-kb yeast artificial chromosome (YAC) that contains a functional copy of the human hypoxanthine phosphoribosyltransferase (HPRT) gene has been isolated. This YAC, yHPRT, and another YAC, yXY837, which contains the 3' end of the HPRT gene, have been mapped with restriction enzymes that cleave human DNA infrequently. The HPRT gene lies near the center of yHPRT. Fusion of yHPRT-containing yeast spheroplasts with mouse L A-9 cells, which are HPRT-negative, gives rise to HPRT-positive colonies. These colonies contain the human HPRT gene and express human HPRT mRNA. Fusion of yeast with mammalian cells is an efficient way of testing the integrity and functionality of human DNA contained in YACs.
一个含有人类次黄嘌呤磷酸核糖转移酶（HPRT）基因功能拷贝的680kb酵母人工染色体（YAC）已被分离出来。这个YAC，即yHPRT，以及另一个包含HPRT基因3'端的YAC，yXY837，已用切割人类DNA频率较低的限制性内切酶进行了图谱绘制。HPRT基因位于yHPRT的中心附近。将含有yHPRT的酵母原生质体与HPRT阴性的小鼠LA-9细胞融合，可产生HPRT阳性菌落。这些菌落含有人类HPRT基因并表达人类HPRT mRNA。酵母与哺乳动物细胞的融合是测试YAC中所含人类DNA完整性和功能的有效方法。
