中国人SRY基因的分离、克隆和核苷酸序列分析

睾丸决定因子基因(Testis-determining factor,TDF)位于Y染色体短臂上,它的表达产物诱导睾丸组织的发生。SRY基因(Sex-determining Region of the Y)位于Y染色体的性别决定区内,许多特征显示SRY就是TDF。我们选用与SRY基因相应的引物,用PCR技术对正常人男女各10例的基因组DNA进行扩增。将特异扩增的男性SRY基因片段重组到质粒PUC12中,得到含有中国人SRY基因片段的克隆,命名为PSY-1、PSY-2。用~(32)p]标记重组质粒中的SRY基因片段作探针,与PCR结果进行Southern杂交,男性样品均显示特异杂交带,女性样品为阴性。用末端终止法测定克隆的SRY基因片段的全部核苷酸序列为299bp,含有SRY基因中高度保守及功能特异性区域的240bp,我们对此进行了讨论。

Isolating, Cloning and Sequencing of the SRY Gene from a Chinese Individual

The testis-determining factor gene (TDF) lies on the Y chromosome and is responsible for initiating male sex detemination. SRY is a gene located in the sex determining region of the human Y chromosome that has many of the properties expected for TDF, The SRY gene from peripheral blood lymphocytes of normal population was amplified by PCR. The specific amplified DNA fragment from a Chinese individual was cloned into vector plasmid pUC_(12). The clone (termed pSY-1 and pSY-2) that contained the SRY gene fragment was obtained. When southern hybridization was performed with amplified DNA fragments using recombinanted SRY gene fragment as a probe, positive results were shown in all males But not in any female. The complete nucleotide sequence of 299 bp of the SRY gene fragment was determined by the dideoxynucleotide termination sequencing method. It contains all 240 bp which is the highly conserved motif and Y specific among the human SRY gene.