The strategy of isolating the band-specific expression fragments from a probe pool generated by humanchromosome microdissection was reported. A cliromosome 14q24.3 band-specific single copy DNA pool was constructed ba...The strategy of isolating the band-specific expression fragments from a probe pool generated by humanchromosome microdissection was reported. A cliromosome 14q24.3 band-specific single copy DNA pool was constructed based on this probe pool. Using total DNA of the pool as probe to hybridize the human marrow cDNA library, 68 primary positive clones were selected from 5 ×105 cDNA clones. Among these primary clones, 32 secondary clones were obtained after second-round screening and designed as cFD14-1～32. Finally, 24 band-specific expression fragments were identified from these 32 positive clones by DNA hybridization. Those band-specific clones can hybridize to both 14q24.3 DNA and human genomic DNA but cann’t hybridize to 17q11～12 DNA. Partial sequences of 13 fragments of them were sequenced and identified as novel cDNA sequences , and these sequences were proved to have some homology with known genes in NCBI database. Analysis of expression spectrum of cFD14-1 suggested that the cDNA fragments thus obtained should be used to isolate the genes can not been cloned in 14q24.3region.展开更多
文摘The strategy of isolating the band-specific expression fragments from a probe pool generated by humanchromosome microdissection was reported. A cliromosome 14q24.3 band-specific single copy DNA pool was constructed based on this probe pool. Using total DNA of the pool as probe to hybridize the human marrow cDNA library, 68 primary positive clones were selected from 5 ×105 cDNA clones. Among these primary clones, 32 secondary clones were obtained after second-round screening and designed as cFD14-1～32. Finally, 24 band-specific expression fragments were identified from these 32 positive clones by DNA hybridization. Those band-specific clones can hybridize to both 14q24.3 DNA and human genomic DNA but cann’t hybridize to 17q11～12 DNA. Partial sequences of 13 fragments of them were sequenced and identified as novel cDNA sequences , and these sequences were proved to have some homology with known genes in NCBI database. Analysis of expression spectrum of cFD14-1 suggested that the cDNA fragments thus obtained should be used to isolate the genes can not been cloned in 14q24.3region.
