Epplen J T, McCarrey J R, Sutou S, Ohno S
Proc Natl Acad Sci U S A. 1982 Jun;79(12):3798-802. doi: 10.1073/pnas.79.12.3798.
A 2.5-kilobase fragment of a sex-specific satellite DNA from the Colubrid snake species Elaphe radiata has been cloned, and its sequence has been determined. It contains 26 and 12 copies, respectively, of two base quadruplets, G-A-T-A and G-A-C-A, as its sole highly repetitious elements. Southern hybridization experiments with genomic DNA of the chicken, the mouse, and man indicated male sex-specific conservation of at least parts of this cloned DNA. In situ hybridization experiments with metaphase chromosomes of the mouse showed that elements that can cross-hybridize with parts of the cloned snake DNA are concentrated in the pericentric region of the Y chromosome. In blot hybridization experiments with liver poly(A)+ polysomal RNAs of male and female mice, a probe consisting of the first 1,224 bases of the cloned snake DNA singled out a male-specific RNA of 1,250-1,400 bases. Inasmuch as the proximal end of this probe contained an open reading frame (44 consecutive amino acid-specifying codons), the male-specific putative mRNA so detected may specify H-Y antigen. By contrast, a probe consisting of bases 1,480-1,906, containing the simple repeats of the quadruplets, singled out a shorter (approximately 1,000-base) RNA from males and females alike. Although this RNA is poly(A)+, we have yet to establish its attachment to ribosomes.
已克隆出游蛇科物种辐射锦蛇的一种性别特异性卫星DNA的2.5千碱基片段,并测定了其序列。它分别含有26个和12个碱基四联体G-A-T-A和G-A-C-A的拷贝,作为其唯一的高度重复元件。用鸡、小鼠和人的基因组DNA进行的Southern杂交实验表明,该克隆DNA的至少部分存在雄性性别特异性保守。用小鼠中期染色体进行的原位杂交实验表明,能与克隆蛇DNA的部分发生交叉杂交的元件集中在Y染色体的着丝粒周围区域。在用雄性和雌性小鼠肝脏的多聚腺苷酸加尾多聚体RNA进行的印迹杂交实验中,由克隆蛇DNA的前1224个碱基组成的探针筛选出一种1250 - 1400个碱基的雄性特异性RNA。由于该探针的近端包含一个开放阅读框(44个连续的氨基酸编码密码子),如此检测到的雄性特异性推定mRNA可能编码H-Y抗原。相比之下,由1480 - 1906碱基组成的探针,包含四联体的简单重复序列,从雄性和雌性中都筛选出一种较短的(约1000个碱基)RNA。尽管这种RNA是多聚腺苷酸加尾的,但我们尚未确定它与核糖体的附着情况。