Tokino T, Takahashi E, Mori M, Tanigami A, Glaser T, Park J W, Jones C, Hori T, Nakamura Y
Division of Biochemistry, Cancer Institute, Tokyo, Japan.
Am J Hum Genet. 1991 Feb;48(2):258-68.
To obtain new RFLP markers on human chromosome 11 for a high-resolution map, we constructed a cosmid library from a Chinese hamster x human somatic hybrid cell line that retains only human chromosome 11 in a Chinese hamster genomic background. A total of 3,500 cosmids were isolated by colony hybridization with labeled human genomic DNA. DNA was prepared from 130 of these cosmid clones and examined for RFLP. In 62 of them, polymorphism was detected with one or more enzymes; four RFLPs were VNTR systems. All polymorphic clones were assigned to one of 22 intervals obtained by mapping on a deletion panel of 15 somatic hybrid cell lines containing parts of chromosome 11; 11 clones were finely mapped by in situ hybridization. Although RFLP markers were scattered on the whole chromosome, they were found predominantly in the regions of R-banding. These DNA markers will contribute to fine mapping of genes causing inherited disorders and tumor-suppressor genes that reside on chromosome 11. Furthermore, as one-third of the cosmid clones revealed a band or bands in Chinese hamster DNA, indicating sequence conservation, this subset of clones may be useful for isolating biologically important genes on chromosome 11.
为了在人类11号染色体上获得新的限制性片段长度多态性(RFLP)标记以构建高分辨率图谱,我们从一个中国仓鼠与人类体细胞杂交细胞系构建了一个黏粒文库,该杂交细胞系在仓鼠基因组背景下仅保留人类11号染色体。通过用标记的人类基因组DNA进行菌落杂交,共分离出3500个黏粒。从其中130个黏粒克隆中制备DNA并检测RFLP。其中62个克隆用一种或多种酶检测到多态性;4个RFLP是可变数目串联重复序列(VNTR)系统。所有多态性克隆都被定位到通过在包含11号染色体部分片段的15个体细胞杂交细胞系的缺失图谱上进行定位而获得的22个区间中的一个;11个克隆通过原位杂交进行了精细定位。尽管RFLP标记分散在整个染色体上,但它们主要出现在R带区域。这些DNA标记将有助于对导致遗传性疾病的基因和位于11号染色体上的肿瘤抑制基因进行精细定位。此外,由于三分之一的黏粒克隆在仓鼠DNA中显示出条带,表明序列保守,这部分克隆可能有助于分离11号染色体上具有生物学重要性的基因。