荷斯坦牛随机扩增微卫星多态性指纹图谱中的雄性特异性条带。

Y M Horng, M C Huang
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引用次数: 0

摘要

共采集了65份荷斯坦牛的血液样本进行DNA提取。采用随机扩增多态性DNA (RAPD)技术扩增基因组DNA。随机选取101条引物(OPAA、OPAO、opv、OPC、OPE和OPA-06)进行聚合酶链反应(PCR)。PCR产物经琼脂糖凝胶电泳分离后,转移到尼龙膜上,与α - 32p标记的微卫星探针杂交,进行随机扩增微卫星多态性(RAMPO)分析。所有引物都在RAMPO指纹图谱中产生了多种扩增模式,但只有OPE-01 (5'-CCCAAGGTCC-3')产生了男性特异性的强信号。65例中,24例男性样本的RAMPO指纹存在特异带,41例女性样本的RAMPO指纹不存在特异带。因此,这种特殊的带是雄性牛所特有的。
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Male-specific band in random amplified microsatellite polymorphism fingerprints of Holstein cattle.

A total of 65 blood samples collected from Holstein cattle were employed for DNA extraction. Genomic DNA were amplified by means of random amplified polymorphic DNA (RAPD). One hundred and one random primers (Operon kits OPAA, OPAO, OPAV, OPC, OPE and OPA-06) were used for polymerase chain reactions (PCR). The PCR products were size fractionated by means of electrophoresis in agarose gel, transferred to a nylon membrane, and then hybridized with a alpha-32P-labeled microsatellite probe for random amplified microsatellite polymorphism (RAMPO) analysis. All the primers produced many amplification patterns in RAMPO fingerprints, but only OPE-01 (5'-CCCAAGGTCC-3') yielded a male-specific strong signal. Among the 65 cases, the specific band could be observed in the RAMPO fingerprints of the 24 male samples but not in the 41 female samples. Therefore, this specific band is peculiar to male cattle.

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