MacLaughlin J, Terner C
Biochem J. 1973 Aug;133(4):635-9. doi: 10.1042/bj1330635.
Spermatozoa from the cauda of the epididymis of the hamster and rat were incubated with [5-(3)H]uridine and glucose. By using a procedure avoiding bacterial and other cellular contamination, sonic extracts were prepared and digested with deoxyribonuclease and Pronase. Radioactive RNA of high molecular weight was isolated by two methods: (a) gel filtration on Sephadex G-75 columns and (b) polyacrylamide-gel electrophoresis in which it migrated in the region of 28S and 23S RNA markers. The macromolecules were alkali-labile and hydrolysed by ribonuclease. From (3)H radioactivity and E(260) of the isolated RNA the rate of incorporation of uridine into RNA of spermatozoa was calculated to be 0.1-0.5nmol/h per mg of RNA.
将来自仓鼠和大鼠附睾尾部的精子与[5-(3)H]尿苷和葡萄糖一起孵育。通过采用避免细菌和其他细胞污染的方法,制备了超声提取物,并用脱氧核糖核酸酶和链霉蛋白酶进行消化。通过两种方法分离出高分子量的放射性RNA:(a) 在Sephadex G-75柱上进行凝胶过滤,以及(b) 聚丙烯酰胺凝胶电泳,在该电泳中它迁移到28S和23S RNA标志物区域。这些大分子对碱不稳定,并被核糖核酸酶水解。根据分离出的RNA的(3)H放射性和E(260),计算出精子RNA中尿苷掺入率为每毫克RNA 0.1 - 0.5nmol/h。