Yasuda J, Navarro J M, Malkhosyan S, Velazquez A, Arribas R, Sekiya T, Perucho M
Oncogene Division, National Cancer Center Research Institute, Tsukiji Chuo-ku, 104 Tokyo, 5-1-1, Japan.
Genomics. 1996 May 15;34(1):1-8. doi: 10.1006/geno.1996.0235.
We have developed a technique for the simultaneous chromosomal assignment of multiple human DNA sequences from DNA fingerprints obtained by the arbitrarily primed polymerase chain reaction (AP-PCR). Radioactively labeled human AP-PCR products are hybridized to DNA fingerprints generated with the same arbitrary primer from human/rodent monochromosome cell hybrids after electroblotting to a nylon membrane. Human-specific hybridization bands in the human/rodent fingerprints unambiguously determine their chromosome of origin. We named this method simultaneous hybridization of arbitrarily primed PCR DNA fingerprinting products (SHARP). Using this approach, we determined the chromosomal origins of most major bands of human AP-PCR fingerprints obtained with two arbitrary primers. Altogether, the chromosomal localization of near 50 DNA fragments, comprehensive of all human chromosomes except chromosomes 21 and Y, was achieved in this simple manner. Chromosome assignment of fingerprint bands is essential for molecular karyotyping of cancer by AP-PCR DNA fingerprinting. The SHARP method provides a convenient and powerful tool for this purpose.
我们已经开发出一种技术,可从通过任意引物聚合酶链反应(AP-PCR)获得的DNA指纹中同时对多个人类DNA序列进行染色体定位。放射性标记的人类AP-PCR产物在电印迹到尼龙膜后,与来自人类/啮齿动物单染色体细胞杂种的相同任意引物产生的DNA指纹杂交。人类/啮齿动物指纹中的人类特异性杂交带明确地确定了它们的起源染色体。我们将此方法命名为任意引物PCR DNA指纹产物的同时杂交(SHARP)。使用这种方法,我们确定了用两种任意引物获得的人类AP-PCR指纹的大多数主要条带的染色体起源。通过这种简单的方式,总共实现了近50个DNA片段的染色体定位,涵盖了除21号和Y染色体之外的所有人类染色体。通过AP-PCR DNA指纹对癌症进行分子核型分析时,指纹条带的染色体定位至关重要。SHARP方法为此提供了一种方便且强大的工具。