Bray P, Lichter P, Thiesen H J, Ward D C, Dawid I B
Laboratory of Molecular Genetics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892.
Proc Natl Acad Sci U S A. 1991 Nov 1;88(21):9563-7. doi: 10.1073/pnas.88.21.9563.
The zinc finger motif, exemplified by a segment of the Drosophila gap gene Krüppel, is a nucleic acid-binding domain present in many transcription factors. To investigate the gene family encoding this motif in the human genome, a placental genomic library was screened at moderate stringency with a degenerate oligodeoxynucleotide probe designed to hybridize to the His/Cys (H/C) link region between adjoining zinc fingers. Over 200 phage clones were obtained and are being sorted into groups by partial sequencing, cross-hybridization with oligodeoxynucleotide probes, and PCR amplification. Further, the genomic clones were cross-hybridized with a set of 30 zinc finger-encoding cDNAs (Kox1-Kox30) isolated from a human T-cell cDNA library. Four cDNAs (Kox4, Kox7, Kox12, and Kox15) were identified that match one or more genomic clones; these matches were confirmed by nucleotide sequence analysis. One or more clones from each locus were mapped onto human metaphase chromosomes by chromosomal in situ suppression hybridization with fluorescent probe detection. We mapped ZNF7/Kox4 to chromosome 8qter, ZNF19/Kox12 to 16q22, ZNF22/Kox15 to 10q11, and ZNF44/Kox7 to 16p11. The results of these analyses support the conclusion that the human genome contains many, probably several hundred, zinc finger genes with consensus H/C link regions.
以果蝇裂隙基因Krüppel的一段序列为代表的锌指基序,是许多转录因子中存在的一种核酸结合结构域。为了研究人类基因组中编码该基序的基因家族,用一种简并寡脱氧核苷酸探针以中等严格度筛选胎盘基因组文库,该探针设计用于与相邻锌指之间的His/Cys(H/C)连接区域杂交。获得了200多个噬菌体克隆,并通过部分测序、与寡脱氧核苷酸探针的交叉杂交和PCR扩增将它们分类成组。此外,将基因组克隆与从人T细胞cDNA文库中分离的一组30个编码锌指的cDNA(Kox1 - Kox30)进行交叉杂交。鉴定出四个与一个或多个基因组克隆匹配的cDNA(Kox4、Kox7、Kox12和Kox15);这些匹配通过核苷酸序列分析得到证实。通过用荧光探针检测的染色体原位抑制杂交,将每个基因座的一个或多个克隆定位到人类中期染色体上。我们将ZNF7/Kox4定位到8号染色体长臂末端,ZNF19/Kox12定位到16号染色体q22,ZNF22/Kox15定位到10号染色体q11,ZNF44/Kox7定位到16号染色体p11。这些分析结果支持这样的结论:人类基因组包含许多,可能有数百个,具有一致H/C连接区域的锌指基因。