Yen C H, Elliott R W
Department of Molecular and Cellular Biology, Roswell Park Cancer Institute, New York State Department of Health, Buffalo 14263, USA.
Genomics. 1997 Jan 1;39(1):19-29. doi: 10.1006/geno.1996.4462.
A library, containing M. spretus DNA in a half-YAC vector, was made and screened for clones hybridizing with an oligomer of the telomere hexamer TTAGGG. FISH to metaphase spreads of spleen cells showed hybridization of clone YTY3 to the distal ends of both X and Y chromosomes, consistent with localization to the pseudoautosomal region (PAR). Recombinational mapping in the BXD RI strains and an interspecies backcross, using a plasmid subclone and PCR primers from YTY3, showed linkage to distal Chr X. The restriction map of the YAC contains three NotI sites. Sequences similar to Mov15 lie close to the vector end of the clone, while the other end contains a telomere array that appears to be interstitial within the PAR, suggesting that the insert represents a proximal portion of the PAR. Sequences homologous to the clone are also present at subtelomeric regions of autosomes 4, 9, and 13.
构建了一个包含斯氏小家鼠DNA的半YAC载体文库,并筛选与端粒六聚体TTAGGG的寡聚物杂交的克隆。对脾细胞中期染色体铺展进行荧光原位杂交(FISH)显示,克隆YTY3与X和Y染色体的远端杂交,这与定位到假常染色体区域(PAR)一致。在BXD RI品系和种间回交中进行重组作图,使用来自YTY3的质粒亚克隆和PCR引物,显示与远端X染色体连锁。YAC的限制酶切图谱包含三个NotI位点。与Mov15相似的序列靠近克隆的载体末端,而另一端包含一个端粒阵列,该端粒阵列似乎位于PAR内间质位置,表明插入片段代表PAR的近端部分。与该克隆同源的序列也存在于4号、9号和13号常染色体的亚端粒区域。