Itoh Y, Ogawa A, Murata K, Hosoda T, Mizuno S
Department of Applied Biological Chemistry, Faculty of Agriculture, Tohoku University, Sendai, Japan.
Genes Genet Syst. 1997 Feb;72(1):51-6. doi: 10.1266/ggs.72.51.
Southern blotting of HindIII-digested genomic DNAs from the female and male Oriental white stork (Ciconia boyciana) with the EE0.6 probe cloned from the W chromosome of chicken (Gallus domesticus) produced a 3.0-kb W chromosome-derived band specific to the female and a 3.5-kb Z chromosome-derived band common to both sexes. These two genomic fragments were cloned and the counterpart sequences to that of EE0.6 in these fragments, XH0.6 and XH0.6RSM (XH0.6-related sequence in the male), respectively, were subcloned. Nucleotide sequences of XH0.6 and XH0.6RSM showed 92% identity. PCR using a set of primer sequences from XH0.6, which differed several nucleotides from those in XH0.6RSM, amplified an about 300-bp genomic sequence only from the female C. boyciana. This method was applied successfully to identify the sex of individual young birds of C. boyciana, an endangered special natural monument in Japan and whose sexes are unidentifiable from their external morphology.
用从鸡（家鸡）W染色体克隆的EE0.6探针，对雌性和雄性东方白鹳（白鹳）经HindIII消化的基因组DNA进行Southern印迹分析，结果显示雌性有一条3.0 kb源自W染色体的特异性条带，而两性都有一条3.5 kb源自Z染色体的共同条带。克隆了这两个基因组片段，并分别亚克隆了这些片段中与EE0.6对应序列的XH0.6和XH0.6RSM（雄性中与XH0.6相关的序列）。XH0.6和XH0.6RSM的核苷酸序列显示出92%的同一性。使用一组来自XH0.6的引物序列进行PCR，该序列与XH0.6RSM的序列相差几个核苷酸，结果仅从雌性白鹳中扩增出一条约300 bp的基因组序列。该方法成功应用于鉴定日本濒危特别天然纪念物白鹳幼鸟个体的性别，其性别从外部形态无法识别。
