摘要
运用染色体显微分离技术 ,从鹌鹑有丝分裂相中分离了Z染色体 ,经DOP PCR扩增后 ,将扩增产物克隆到pBluescript质粒中 ,构建了鹌鹑Z染色体的特异DNA文库。以该文库中分离的插入DNA作为探针 ,对鹌鹑染色体进行了描绘。结果在鹌鹑的分裂相中 ,Z染色体被强烈的特异荧光信号完全覆盖 ,而在其他染色体上基本没有杂交信号。表明该文库中的插入DNA对鹌鹑Z染色体是特异的 ,可以用来制备性染色体的描绘探针 ,在其他相关物种中检测Z染色体的保守同线群。
A simple method was used to adapt a standard light microscope for the collection of quail Z chromosomes from mitotic metaphase spreads. The microisolated chromosomes were subjected to proteinase K treatment in a collection drop to release DNA, which was then amplified using a degenerate oligonucleotide primed PCR(DOP PCR) strategy. Size distributions of the PCR products were analyzed by agarose gel electrophoresis, and smears of DNA revealed that ranged in size from 200～1 400bp, without any evidence of preferential amplification. The second round PCR products were cloned into pBluescript plasmids to construct a Z chromosome specific DNA library. The size range of the cloned inserts was 200～1 400bp. Using inserted fragments from the library as probes, chromosome painting was performed on quail chromosomes. The results showed that Z chromosomes of quail were completely covered by strong signals and there were little signals on other chromosomes. It was indicated that inserted DNA of the library was specific to the Z chromosome of quail. The library can be used as chromosome painting probe to detect conserved syntenic groups on the chromosomes of other related species and study mechanisms of sex chromosomes evolution in birds.
基金
国家自然科学基金 (No .3 9970 40 6)
教育部博士学科点专项科研基金 (No .980 4862 8)资助
