Lindgren G, Breen M, Godard S, Bowling A, Murray J, Scavone M, Skow L, Sandberg K, Guérin G, Binns M, Ellegren H
Department of Evolutionary Biology, Norbyvägen, Uppsala University, Sweden.
Chromosome Res. 2001;9(1):53-9. doi: 10.1023/a:1026743700819.
We report fluorescence in-situ hybridization (FISH) and somatic cell hybrid mapping data for 13 different horse genes (ANP, CD2, CLU, CRISP3, CYP17, FGG, IL1RN, IL10, MMP13, PRM1, PTGS2, TNFA and TP53). Primers for PCR amplification of intronic or untranslated regions were designed from horse-specific DNA or mRNA sequences in GenBank. Two different horse bacterial artificial chromosome (BAC) libraries were screened with PCR for clones containing these 13 Type I loci, nine of which were found in the libraries. BAC clones were used as probes in dual colour FISH to confirm their precise chromosomal origin. The remaining four genes were mapped in a somatic cell hybrid panel. All chromosomal assignments except one were in agreement with human-horse ZOO-FISH data and revealed new and more detailed information on the equine comparative map. CLU was mapped by synteny to ECA2 while human-horse ZOO-FISH data predicted that CLU would be located on ECA9. The assignment of IL1RN permitted analysis of gene order conservation between HSA2 and ECA15, which identified that an event of inversion had occurred during the evolution of these two homologous chromosomes.
我们报告了13种不同马基因（心钠素、CD2、CLU、CRISP3、CYP17、纤维蛋白原γ链、白细胞介素1受体拮抗剂、白细胞介素10、基质金属蛋白酶13、精子蛋白1、前列腺素内过氧化物合酶2、肿瘤坏死因子α和p53）的荧光原位杂交（FISH）和体细胞杂交定位数据。用于内含子或非翻译区PCR扩增的引物是根据GenBank中马特异性DNA或mRNA序列设计的。用PCR筛选了两个不同的马细菌人工染色体（BAC）文库，以寻找含有这13个I型位点的克隆，其中9个在文库中被发现。BAC克隆用作双色FISH的探针，以确认其精确的染色体来源。其余4个基因在体细胞杂交板中进行定位。除一个外所有染色体定位均与人类-马全基因组荧光原位杂交（ZOO-FISH）数据一致，并揭示了马比较图谱上新的和更详细的信息。CLU通过同线性定位到马1号染色体（ECA2），而人类-马ZOO-FISH数据预测CLU将位于马9号染色体（ECA9）上。白细胞介素1受体拮抗剂（IL1RN）的定位允许分析人类2号染色体（HSA2）和马15号染色体（ECA15）之间的基因顺序保守性，这表明在这两条同源染色体的进化过程中发生了一次倒位事件。
