作　　者： ; ; ; ; ;

机构地区： 江西农业大学

出　　处： 《农业生物技术学报》 2005年第6期759-762,共4页

摘　　要： 牛牙釉蛋白(AML)基因定位于牛X染色体和Y染色体上的同源区域。实验根据牛X、Y染色体AML基因第五内含子上序列同源性只有45.1%以及X、Y染色体该区段之间存在多处碱基缺失的特点,合成了1对性别特异性引物。该引物经扩增后母牛只得到1条467bp的来自X染色体的扩增条带,而公牛能得到1条源自X染色体的467bp片段的扩增条带和1条源自Y染色体的341bp的扩增条带。经基因组DNA验证,该引物的公母鉴定准确率为100%,同时该对引物具有高度的牛特异性。定量分析AML基因引物的反应灵敏度,发现1次PCR(30个循环)能扩增出0.5ng的基因组DNA,2次PCR(共50个循环)能扩增出20pg的基因组DNA。将AML基因引物与SRY基因引物的反应灵敏度相比较,结果表明前者的灵敏度比后者的高8～10倍。用AML基因反应体系鉴定了26枚优质胚胎,其中24枚有扩增结果,13枚鉴定为公牛,11枚鉴定为母牛。 Amelogenin（AML） gene locates on the conservative region of the bovine X and Y chromosomes. Based on the low homology （45.1%） between the fifth intron of the bAML-X and bAML-Y gene with multiple deletions,a pair of gender-specific primers were utilized to amplify a 467-bp fragment from the female bovine X-chromosome, and a 467-bp fragment from X-chromosome and a 34 l-bp fragment from the Y-chromosome in male bovine, respectively. Bovine genomic DNA with known gender were used as template, the accuracy and specificity of the primers were confirmed. The sensitivity of AML primers was analysed quantitatively. Amplicons were obtained by single PCR （30 cycles） using 0.5 ng ofgenomic DNA, and by dual PCR （50 cycles） using 20 pg of genomic DNA as template. Comparing with SRY primers, the AML primers have more than 8-10 times sensitivity. Our protocol of embryo sexing was applied to a total of 26 Holstein dairy cattle embryos ,13 were identified as male and the other ones were identified as female.

关 键 词： 基因 牛 胚胎 性别鉴定

领　　域： [农业科学] [农业科学] [农业科学]