牛雄性DNA特异的限制性片段研究

牛基因组DNA经PstI酶解后,以与性别有关的雌性蛇的小卫星DNA组份一BKm序列(p184)为探针进行分子杂交,回收4.4kb牛雄性DNA片段,以pUC_(19)为载体亚克隆,重组质粒经菌落原位杂交和Southern blot分子杂交鉴定。将此雄性DNA片段检测牛基因组DNA,观察到由于性别的不同而出现不一样的杂交模式。实验结果表明,用来自牛雄性DNA特异的限制性片段的重组质粒作探针,Southern blot分子杂交方法可以鉴定牛的性别。     

牛Y染色体DNA富集文库的构建

According to the fundamentals of DNA renatured kinetics, we made a study in which genomic DNA from male cattle was completely digested with Sau 3A, while female DNA was sheared to an average size of 500 bp fragments. The Y derived DNA sequence rich library of bovine has been contracted with pUC19 plasmid as vector and JM109 as host bacteria by reassociation of male and female DNA. Male and female DNA probes, were used. The result of colony hybridization showed that out of 1000 recombinants analysed, 6 were found to be stronger signal with male probe. Analysis of Pvu' I digested DNA fragment revealed that most of the inserts were in the 90-300 bp.     

牛Y染色体DNA富集文库的构建

作者根据DNA复性动力学原理,使牛雄性基因组DNA经Sau3A完全消化,雌性DNA被剪切到分子量平均500bp大小,二者混合重聚反应后,以pUC_(19)为载体,JM109为宿主菌,构建了Y染色体DNA富集文库。分别以雄性和雌性基因组DNA作探针进行菌落原位杂交,结果表明,1000个克隆中约有6个克隆对雄性DNA探针产生强的杂交信号。PvuⅡ酶切重组质粒,大多数质粒有短片段插入,分子量在90～300bp之间。     

牛雄性DNA特异的限制性片段研究

Bkm, which is related to sexes, is a satellite DNA of the female snake. After the genomic DNA of bovine was digested by Pst I, it was hybridized with Bkm sequence probe (p184) and subcloned in the vector pUC19. Recombinants were identified by the colony in situ and Southern blot hybridization. When we determined genomic DNA of bovine using the male DNA fragment, it was observed that the pattern of hybridization was different because of different sexes. The results showed that we could identify the sex of bovine by Southern blot hybridization with recombinants of male DNA special restricted fragment of bovine origin.