Kishino T, Ariga T, Soejima H, Tamura T, Ohta T, Jinno Y, Yonemura S, Sato N, Tsukita S, Tsukita S
Department of Human Genetics, Nagasaki University School of Medicine, Japan.
Cytogenet Cell Genet. 1994;66(3):167-9. doi: 10.1159/000133692.
The human moesin gene (MSN) was mapped to the long arm of the X chromosome. PCR products for the moesin gene cDNA were used as probes to isolate their corresponding cosmid clones. Fluorescence in situ hybridization (FISH) with two of the isolated cosmid probes showed signals at Xq11.2-->q12, whereas four other cosmids showed FISH signals on chromosome 5. Southern blot hybridization, using a PCR product corresponding to the 3' region of the moesin gene cDNA as a probe (probe-3), on one of the two cosmids that produced signals on the X chromosome gave 5.7- and 3.5-kb HindIII fragments. Further Southern hybridization of the DNA from XY, XX, and XXXXX individuals using probe-3 revealed a gene-dose effect of the X chromosome on the size of a 3.5-kb and a 3.0-kb HindIII fragment; in contrast, an invariant 9.8-kb band was present in the DNA of all individuals tested. Sequencing of an exon-intron border revealed that the two cosmids had predicted sequences. These results indicated that the two cosmids contained MSN, and it was consequently assigned to human chromosome region Xq11.2-->q12. These results strongly suggest that MSN may be removed from candidacy for Wiskott-Aldrich syndrome, which has been putatively mapped to Xp11.3-->p11.22.
人类埃兹蛋白基因（MSN）被定位到X染色体长臂。用埃兹蛋白基因cDNA的PCR产物作为探针来分离其相应的黏粒克隆。用两个分离出的黏粒探针进行荧光原位杂交（FISH），结果显示在Xq11.2→q12处有信号，而另外四个黏粒在5号染色体上显示FISH信号。用对应于埃兹蛋白基因cDNA 3'区域的PCR产物作为探针（探针-3）进行Southern印迹杂交，在两个在X染色体上产生信号的黏粒之一上得到了5.7 kb和3.5 kb的HindIII片段。使用探针-3对XY、XX和XXXXX个体的DNA进行进一步的Southern杂交，结果显示X染色体对3.5 kb和3.0 kb的HindIII片段大小存在基因剂量效应；相反，在所有测试个体的DNA中都存在一条恒定的9.8 kb条带。对一个外显子-内含子边界进行测序，结果显示这两个黏粒具有预测序列。这些结果表明这两个黏粒包含MSN，因此将其定位到人类染色体区域Xq11.2→q12。这些结果强烈表明，MSN可能不再是威斯科特-奥尔德里奇综合征候选基因，该综合征此前被推测定位在Xp11.3→p11.22。
