Weier H U, Miller B M, Yu L C, Fuscoe J C
Department of Laboratory Medicine, School of Medicine, University of California, San Francisco 94143-0808.
DNA Seq. 1993;4(1):47-51. doi: 10.3109/10425179309015622.
Hamster chromosome-specific DNA sequences were amplified by primer directed DNA amplification using mixed base oligonucleotides in an arbitrarily primed polymerase chain reaction (AP-PCR) protocol. The template DNA was comprised of approximately 3000 chinese hamster ovary cell (CHO) chromosomes enriched by flow sorting from a human x hamster hybrid cell line. Labeling of the PCR product pool and fluorescence in situ hybridization (FISH) demonstrated preferential binding to the distal long arm of the CHO X chromosome. The PCR products were cloned, labeled by PCR and hybridized to metaphase spreads. Clones containing highly reiterated DNA were identified by FISH and sequenced. Here, we present the sequence and chromosomal location of one of the repeat clones that maps close to the secondary constriction on the long arm of the CHO X chromosome, pCAT2066-24.
使用混合碱基寡核苷酸，通过引物定向DNA扩增，在任意引物聚合酶链反应（AP-PCR）方案中扩增仓鼠染色体特异性DNA序列。模板DNA由大约3000条中国仓鼠卵巢细胞（CHO）染色体组成，这些染色体通过流式分选从人x仓鼠杂交细胞系中富集得到。PCR产物池的标记和荧光原位杂交（FISH）显示其优先结合到CHO X染色体的远端长臂上。PCR产物被克隆，通过PCR进行标记，并与中期染色体铺展进行杂交。通过FISH鉴定出含有高度重复DNA的克隆并进行测序。在此，我们展示了一个重复克隆的序列和染色体定位，该克隆定位于CHO X染色体长臂上靠近次缢痕的位置，即pCAT2066 - 24。
