Zheng L, Saunders R D, Fortini D, della Torre A, Coluzzi M, Glover D M, Kafatos F C
Biological Laboratories, Harvard University, Cambridge, MA 02138.
Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11187-91. doi: 10.1073/pnas.88.24.11187.
We have microdissected divisions of the Anopheles gambiae polytene chromosomes, digested the DNAs with a restriction enzyme, and PCR-amplified the DNA fragments to generate a set of pooled probes, each corresponding to approximately 2% of the mosquito genome. These divisional probes were shown to have high complexity. Except for those derived from near the centromeres, they hybridize specifically with their chromosomal sites of origin. Thus, they can be used to map cloned DNAs by a dot blot procedure, which is much more convenient than in situ hybridization to polytene chromosomes. We discuss additional potential uses of these probes, such as easier isolation of molecular markers and genes, including those that cross-hybridize with clones available from other insects. It is expected that the probes will substantially accelerate molecular genetic analysis of this most important malaria vector.
我们对冈比亚按蚊多线染色体进行了显微切割，用一种限制性酶消化DNA，并通过PCR扩增DNA片段以生成一组混合探针，每个探针大约对应蚊子基因组的2%。这些分区探针显示具有高复杂性。除了那些源自着丝粒附近的探针外，它们与其染色体起源位点特异性杂交。因此，它们可用于通过斑点印迹法对克隆的DNA进行定位，这比多线染色体原位杂交要方便得多。我们讨论了这些探针的其他潜在用途，例如更容易分离分子标记和基因，包括那些与其他昆虫可获得的克隆交叉杂交的标记和基因。预计这些探针将大大加速对这种最重要的疟疾传播媒介的分子遗传分析。
