摘要
本文报道了从显微切割的人染色体区带直接分离区带专一性表达序列的方法和结果。
The strategy of isolating the band-specific expression fragments from the probe pool of human chromosome generated by microdissection was reported in present paper. A chromosome 14q24.3 band-specific single copy DNA library was constructed based on this probe pool. Using this pool DNA as probe to hybridize the human bone marrow cell cDNA libray, 68 primary positive clones were selected from 5 × 105 cDNA clones. Of them 32 clones were got in second-round screening and designed as cFD14-1-32. Finally, 24 bandspecific expression fragments were identified from these 32 positive
clones by analysing the results of DNA hybridization. Those band-specific clones can hybridize to both 14 q 24.3 DNA and human genomic DNA, but have no hybridization signal with 17q11-12 DNA. Partial sequences of 13 fragments of them were sequenced and were identified as novel cDNA sequences as well as have some homology with known genes in NCBI database. Analysis of expression spectrum of cFD 14-1 suggested that the cDNA fragments thus obtained can be used to isolate the genes not yet be cloned in 14 q 24.3 region.
出处
《实验生物学报》
CSCD
1997年第3期241-246,共6页
Acta Biologiae Experimentalis Sinica
基金
上海市科技发展重点项目
国家自然科学基金
关键词
染色体
14q24.3区带
探针
基因表达序列
分离
Probe pool generated by microdissection. Chromosome 14 q 24.3 band. Single copy DNA library of chromosome 14 q 24.3. Expression sequences of genes
