染色体显微切割与DOP-PCR结合对赤麂Sry基因克隆、测序及初步定位  

Cloning, Sequencing and Chromosome Location of Sry Gene of Muntjak munticus vaginalis by DOP-PCR and Microdissection

作　　者：张悦[1] 鲁晓萱[1] 刘宁生[1] 余多慰[2] 单祥年[2] 

机构地区：[1]东南大学医学院生物教研室,南京210009 [2]南京师范大学生命科学院,南京210097

出　　处：《Acta Genetica Sinica》2001年第4期322-326,共5页

基　　金：国家自然科学基金资助!(No.39670393)

摘　　要：应用显微切割技术获得赤麂１号、Ｙ１、Ｙ２染色体，通过ＤＯＰ－ＰＣＲ增加模板ＤＮＡ拷贝数，然后用人的性别决定基因（Ｓｅｘ－ｄｅｔｅｒｍｉｎｉｎｇ Ｒｅｇｉｏｎ ｏｆ ｔｈｅ Ｃｈｒｏｍｏｓｏｍｅ Ｙ， ＳＲＹ）中ＨＭＧ框内设计１对引物，对ＤＯＰ－ＰＣＲ产物进行扩增。在雄性赤麂Ｙ２染色体ＤＯＰ－ＰＣＲ产物中扩增出与人ＳＲＹ基因同源的Ｓｒｙ基因片段，克隆、测序，首次在分子水平上证明赤麂Ｙ２染色体是真正的Ｙ染色体，同时对赤麂Ｓｒｙ基因进行了初步定位。We isolated 1, Y1, Y2 chromosomes of the male M.m vaginalis by microdissection and amplified the DNA copies by DOP-PCR. Using the DOP-PCR products of 1, Y1, Y2 chromosomes of M.m vaginalis as template respectively and the primers designed within human SRY HMG box, the Sry gene of the male M.m vaginalis was amplified, and it was cloned and sequenced. It is proved that Y2 is the real sex chromosome in the male M.m vaginalis at molecular level for the first time, and Sry was localized on Y2 chromosome.

关 键 词：染色体 显微切割 DOP-PCR SRY基因 赤麂 性染色体 基因定位 

分 类 号：Q953[生物学—动物学] Q959.8