Mann G B, Fowler K J, Grail D, Dunn A R
Melbourne Tumour Biology Branch, Ludwig Institute for Cancer Research, PO Royal Melbourne Hospital, Victoria, Australia.
J Reprod Fertil. 1993 Nov;99(2):505-12. doi: 10.1530/jrf.0.0990505.
In this study a rapid, simple and inexpensive procedure is described which allows potential germ-line male mice to be identified with confidence. Spermatozoa recovered by uterine washing following mating with normal female mice was analysed in two ways. First, the patterns of expression of the different isoforms of glucose phosphate isomerase were determined. Since the glucose phosphate isomerase isoforms expressed in embryo stem (ES) cell lines are frequently different from those associated with the host blastocyst, it is possible to determine the proportion of spermatozoa produced by an individual animal that was of ES cell or host-blastocyst origin. Second, DNA of spermatozoa was subjected to polymerase chain reaction (PCR) analysis using primers with specificity for the targeted mutation in the ES cells. The PCR analysis was particularly valuable in identifying germ cell chimaeras in which the contribution of ES-derived spermatozoa was significantly less than that specified by the host blastocyst.
在本研究中，描述了一种快速、简单且廉价的方法，该方法能够可靠地鉴定出具有潜在种系的雄性小鼠。与正常雌性小鼠交配后，通过子宫冲洗回收的精子以两种方式进行分析。首先，测定磷酸葡萄糖异构酶不同同工型的表达模式。由于胚胎干细胞（ES）系中表达的磷酸葡萄糖异构酶同工型通常与宿主囊胚相关的同工型不同，因此可以确定由ES细胞或宿主囊胚来源的单个动物产生的精子比例。其次，使用对ES细胞中靶向突变具有特异性的引物对精子DNA进行聚合酶链反应（PCR）分析。PCR分析在鉴定生殖细胞嵌合体方面特别有价值，其中ES来源的精子的贡献明显小于宿主囊胚所规定的贡献。
