Tynan K, Hoffman S M, Tsujimoto S, Brandriff B, Gordon L, Carrano A V, Mohrenweiser H W
Human Genome Center, Lawrence Livermore National Laboratory, Livermore, California 94550.
Genomics. 1993 Aug;17(2):316-23. doi: 10.1006/geno.1993.1327.
The D19S11 locus has been previously described as consisting of a complex set of six nonallelic polymorphic sites detected with a combination of four restriction enzymes and three probes that were subcloned from a single cosmid. These probes also hybridized to additional nonvariant fragments on Southern blots of human genomic DNA. In the course of establishing a contig map of human chromosome 19, a set of cosmids that were positive for at least one of the probes defining this locus was identified. These cosmids, along with additional cosmids, were assembled using a combination of strategies, including fluorescence in situ hybridization studies using G1 interphase nuclei and sperm pronuclei as chromatin targets, into a single overlapping set of cosmids that spans approximately 650 kb. Cosmids that are positive for the MEL gene probe are localized at the centromeric end of the spanning path, with some cosmids being positive for both the MEL gene probe and one of the D19S11 probes. The EcoRI fragments with homology to the various probes have been identified; some cosmids have homology to all three D19S11 probes. The positions for five of the six polymorphic sites were localized within a 40-kb region, with four sites within 15 kb.
D19S11基因座先前已被描述为由一组复杂的六个非等位多态性位点组成，这些位点是通过四种限制性内切酶和三个从单个黏粒亚克隆的探针组合检测到的。这些探针在人类基因组DNA的Southern印迹上也与其他非变异片段杂交。在构建人类19号染色体重叠群图谱的过程中，鉴定出一组对定义该基因座的至少一种探针呈阳性的黏粒。这些黏粒与其他黏粒一起，使用多种策略进行组装，包括以G1间期核和精子原核作为染色质靶标的荧光原位杂交研究，形成一个跨度约650 kb的单一重叠黏粒集。对MEL基因探针呈阳性的黏粒位于跨越路径的着丝粒末端，一些黏粒对MEL基因探针和D19S11探针之一均呈阳性。已鉴定出与各种探针具有同源性的EcoRI片段；一些黏粒与所有三个D19S11探针均具有同源性。六个多态性位点中的五个位点位于一个40 kb的区域内，其中四个位点位于15 kb范围内。
