从显微切割的猪12号染色体短臂分离新微卫星及应用研究

Using the Guangxi Bama miniature swine, one of local breed of our country, as materials, The microdissection of single swine chromosome 12 was carried out. Swine metaphase spreads were prepared from culture of peripheral blood lymphocytes. Swine chromosome 12 microdissection technology was established first time in China and abroad and the smallest mid-centromere chromosome was dissected successfully. The single chromosome 12 was scraped from metaphase spreads , amplified respectively by linker-adaptor (LA-PCR) PCR and degenerate oligunucleotide-primed-PCR (DOP-PCR) successfully. The utility of swine genomic DNA probe for Southern blot was demonstrated and confirmed that The amplified product was verified come frome swine genomic DNA. The microsatellite primers located on chromosome 12 were amplified by PCR reaction and it was verified that the amplification DNAs came from swine chromosome12 by sequenced. The chromosome 12 DNA library was established by cloning the PCR product that can be use to explore new microsatellites and new genes of chromosome 12. The method used here provide valued materials for the obtention of genes and microsatellites.

用本室研究出的方法制备猪（本研究用中国小型猪）二价体标本，在我国首次在猪中用显微切割法获得者2号染色体