Bowcock A, Osborne-Lawrence S, Barnes R, Chakravarti A, Washington S, Dunn C
Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas 75235-9063.
Genomics. 1993 Feb;15(2):376-86. doi: 10.1006/geno.1993.1071.
Twelve polymorphic (CA)n microsatellites were isolated from a flow-sorted chromosome 13 genomic library. These, and two others that have been previously described, were genotyped in 41 families from the CEPH (Centre d'Etude Polymorphisme Humain, Paris), and a primary linkage map with considerable support for order (odds > 10,000:1) was constructed. Two RFLP-based markers, COL4A1 and D13S52, with heterozygosities above 0.67 and an RFLP-based centromeric marker at D13Z1 were included in this map which extends from 13cen to 13q34. The heterozygosity of all of the PCR-based markers is above 60%. The total map spans a genetic distance of 144 cM, extending from D13Z1 to D13S52 with a single maximum intermarker recombination distance of 35 cM. All other intermarker recombination distances are 18 cM or less. Marker order was confirmed by sublocalizing many of the microsatellite containing clones on a panel of rodent-human somatic cell hybrids with deletions and rearrangements of chromosome 13. One spontaneous new mutation for these 14 (CA)n repeat markers was identified from a total of 8006 gametes, giving an overall observed spontaneous mutation rate of 0.00012 per locus per gamete. An integrated map of chromosome 13q was constructed with the microsatellite markers described here and previously genotyped RFLP-based markers. This sex-average map spans 209 cM with an average distance between unique map locations of 4.5 cM; the maximum intermarker distance was 14 cM.
从经流式细胞仪分选的13号染色体基因组文库中分离出12个多态性(CA)n微卫星。将这些微卫星以及另外两个先前已描述的微卫星,在来自CEPH(巴黎人类多态性研究中心)的41个家系中进行基因分型,并构建了一个具有相当可靠顺序支持(优势比>10000:1)的初步连锁图谱。该图谱从13cen延伸至13q34,包含两个基于RFLP的标记COL4A1和D13S52,其杂合度高于0.67,以及一个位于D13Z1的基于RFLP的着丝粒标记。所有基于PCR的标记的杂合度均高于60%。整个图谱跨度为144厘摩的遗传距离,从D13Z1延伸至D13S52,单个标记间最大重组距离为35厘摩。所有其他标记间重组距离均为18厘摩或更小。通过将许多含有微卫星的克隆亚定位到一组具有13号染色体缺失和重排的啮齿动物 - 人类体细胞杂种上,确认了标记顺序。在总共8006个配子中,鉴定出这14个(CA)n重复标记的一个自发新突变,每个位点每个配子的总体观察到的自发突变率为0.00012。利用本文所述的微卫星标记和先前已进行基因分型的基于RFLP的标记构建了13q染色体的整合图谱。这个性别平均图谱跨度为209厘摩,独特图谱位置之间的平均距离为4.5厘摩;最大标记间距离为14厘摩。