Huang Audrey M, Rehm E Jay, Rubin Gerald M
Cold Spring Harb Protoc. 2009 Apr;2009(4):pdb.prot5199. doi: 10.1101/pdb.prot5199.
The Drosophila melanogaster P-transposable element is a powerful and widely used research tool. Sequences flanking the P-element can be recovered and the site of insertion can be mapped to the nucleotide, to connect the genetic and physical maps and facilitate molecular analysis of the gene of interest. The Berkeley Drosophila Genome Project (BDGP) has assembled a well-characterized collection of lethal mutations induced by single P-element insertions generated by a number of laboratories. The genomic DNA sequences adjacent to these insertions have been recovered by either plasmid rescue or inverse polymerase chain reaction (PCR). The combination of a complete genomic DNA sequence and relatively fast and easy molecular methods for mapping P-element insertion sites to the nucleotide enhances the use of P-elements as tools in Drosophila research. This protocol provides detailed procedures for isolating DNA flanking P-element insertions.
黑腹果蝇P转座因子是一种强大且广泛应用的研究工具。P因子两侧的序列能够被回收，并且插入位点能够定位到核苷酸水平，从而连接遗传图谱和物理图谱，便于对感兴趣的基因进行分子分析。伯克利果蝇基因组计划（BDGP）收集了一系列特征明确的致死突变，这些突变由多个实验室产生的单个P因子插入所诱导。与这些插入相邻的基因组DNA序列已通过质粒拯救或反向聚合酶链反应（PCR）回收。完整的基因组DNA序列与相对快速简便的将P因子插入位点定位到核苷酸水平的分子方法相结合，增强了P因子在果蝇研究中作为工具的应用。本方案提供了分离P因子插入侧翼DNA的详细步骤。
