Sato H, Koide T, Sagai T, Ishiguro S I, Tamai M, Saitou N, Shiroishi T
Department of Ophthalmology, Tohoku University School of Medicine, Seiryo-Machi 1-1, Sendai, 980-8574, Japan.
Genomics. 1999 Mar 15;56(3):303-9. doi: 10.1006/geno.1998.5721.
We isolated two new keratin cDNAs by screening a cDNA library constructed from poly(A)+ RNA of the dorsal and abdominal skin of C57BL/10J mice with a probe of human KRT14. Due to its high sequence homology to human keratin 17 cDNA, one full-length cDNA is most likely to be mouse keratin 17 (Krt1-17) cDNA. The other is the putative full-length cDNA of a novel type I keratin gene, designated Krt1-c29. These two keratin genes were mapped to the distal portion of Chromosome 11, where the mouse keratin gene complex-1 (Krt1) is localized. To elucidate the genomic organization of Krt1 in mice, we carried out genetic and physical analyses of Krt1. A large-scale linkage analysis using intersubspecific backcrosses suggested that there are two major clusters in Krt1, one containing Krt1-c29, Krt1-10, and Krt1-12 and the other containing Krt1-14, -15, -17, and -19. Truncation experiments with two yeast artificial chromosome clones containing the two clusters above have revealed that the gene order of Krt1 is centromere-Krt1-c29-Krt1-10-Krt1-12-Krt1-13-K rt1-15-Krt1-19-Krt1-14-K rt1-17-telomere. Finally, we analyzed sequence divergence between the genes belonging to the Krt1 complex. The results clearly indicated that genes are classified into two major groups with respect to phylogenetic relationship. Each group consists of the respective gene cluster demonstrated by genetic and physical analyses in this study, suggesting that the physical organization of the Krt1 complex reflects the evolutionary process of gene duplication of this complex.
我们用人类KRT14探针筛选由C57BL/10J小鼠背部和腹部皮肤的聚腺苷酸加尾RNA构建的cDNA文库，分离出两个新的角蛋白cDNA。由于其与人类角蛋白17 cDNA具有高度序列同源性，一个全长cDNA很可能是小鼠角蛋白17（Krt1-17）cDNA。另一个是新型I型角蛋白基因的推定全长cDNA，命名为Krt1-c29。这两个角蛋白基因被定位到11号染色体的远端，小鼠角蛋白基因复合体-1（Krt1）位于该区域。为了阐明小鼠中Krt1的基因组组织，我们对Krt1进行了遗传和物理分析。使用种间回交的大规模连锁分析表明，Krt1中有两个主要簇，一个包含Krt1-c29、Krt1-10和Krt1-12，另一个包含Krt1-14、-15、-17和-19。对包含上述两个簇的两个酵母人工染色体克隆进行截短实验，揭示了Krt1的基因顺序为着丝粒-Krt1-c29-Krt1-10-Krt1-12-Krt1-13-Krt1-15-Krt1-19-Krt1-14-Krt1-17-端粒。最后，我们分析了属于Krt1复合体的基因之间的序列差异。结果清楚地表明，就系统发育关系而言，基因被分为两个主要组。每组由本研究中通过遗传和物理分析证明的各自基因簇组成，这表明Krt1复合体的物理组织反映了该复合体基因复制的进化过程。
