遗传 ›› 2001, Vol. 23 ›› Issue (2): 114-118.

• 论文 • 上一篇    下一篇

黑麂Y染色体的鉴别和Sry基因的克隆及定位

王毅1; 单祥年1;张悦1;刘宁生1;鲁晓萱1;佴文惠2;王金焕2;陈玉泽2;杨凤堂2;胡均3;张文兴3;徐春茂3 WANG Yi1;SHAN Xiang-nian1;ZHANG Yue1;LIU Ning-sheng1;LU Xiao-xuan1;NIE Wen-hui2;WANG Jin-huan2;CHEN Yu-ze2;YANG Feng-tang2;HU Jun3;ZHANG Wen-xing3;XU Chun-mao3   

  1. 1.东南大学医学院生物教研室,南京 210009; 2.中国科学院昆明动物所,昆明 650223; 3.安徽省休宁野生动物园,休宁 245600 1.Depantment of Biology,Southeast Medical College,Nanjing 210009,China; 2.Kunming institute of Zoology of Chinese Academy of science,Kunming 650223,China; 3.The Zoo of Xiouning of the Anhui Province, Xiouning 245600,China
  • 收稿日期:1900-01-01 出版日期:2001-04-10 发布日期:2001-04-10

The Study of the Sry Gene of Muntiacus crinifrons

  • Received:1900-01-01 Online:2001-04-10 Published:2001-04-10

摘要: 以流式细胞仪分离小麂(Muntiacus reevesi)Y染色体和黑麂(Muntiacus crinifrons)Y1,Y2,X+4和1号染色体,利用DOP-PCR技术富集了分离的各单条染色体。然后,将小麂的Y染色体的DOP-PCR产物经Cy3标记后直接作为涂染探针,应用染色体涂染技术与雌雄黑麂的核型标本进行杂交,确认了黑麂真正的Y染色体为Y2染色体。再以黑麂的Y1,Y2,X+4和1号染色体的DOP-PCR产物为模板,用人的特异性的SRY(sex determining region of the Y chromosome)基因引物对其进行扩增,结果表明黑麂只有Y2染色体出现了SRY扩增片段。然后扩增产物克隆和测序,比较它与人的同源性,初步把黑麂的Sry基因定位在Y2染色体上。最后提取雄性黑麂的基因组DNA,并用同一对引物对其进行扩增,亦得到Sry基因的片段,对此扩增片段进行克隆,测序,结果表明其与Y2染色体得到的Sry基因片段完全一样,与人SRY基因的同源性均为83%。
Abstract:The single Y chromosome of Muntiacus reevesi and Y1,Y2 ,X+4,1 chromosome of Muntiacus crinifrons were obtained by flow-sorting ,then they were amplified through DOP-PCR . After that, the metaphase karyotype of Muntiacus crinifrons were painted by using the product of the DOP-PCR of the Y chromosome of Muntiacus reevesi as a special probe and the result showed that Y2 chromosome was the real Y chromosome of Muntiacus crinifrons. Secondly the product of the DOP-PCR of Y1,Y2,X+4,1 chromosome of Muntiacus crinifrons were used as the templates of the next amplification using the special primer devised according to the human SRY gene .One band was obtained only from Y2 chromosome, then it was cloned to the T-vector and sequenced. The Sry gene sequence of Muntiacus crinifrons was acquired and the conclution was that there are 83% homology between the human and Muntiacus crinifrons. It was testified that in all mammal Sry gene is consertive. On the other side the Sry gene was located to the Y2 chromosome of the Muntiacus crinifrons.

关键词: DOP-PCR, Sry基因, 黑麂, 染色体涂染, 基因定位 Muntiacus crinifrons