单只不育果蝇未知基因的反向PCR

根据ｐ转位子诱变和ｐｏｘｎ基因双杂合子法检测化学感受器数目的变化，筛选到一系列突变个体，其中一个雄蝇腿部表现出不能形成化学感受器的突变表型；遗憾的是该只雄蝇是不良的，为了克隆该基因，研究出一种微量ＤＮＡ反向ＰＣＲ法，该法可用０．５只果蝇扩增出Ｐ转位子插入点相邻的基因组片断，方法简便可靠。

Inverse PCR of Unknown Genes in Single Sterile Fly

We isolated a series of mutants by looking for insertion of a P lacW transposon that show a reduced number of chemosensory bristles when combined in double heterozygote with a deficiency of the poxn gene.One of them showed loss of all chemosensory organs on its legs.Unfortunately,this male fly was sterile .In order to clone this gene,a successful inverse PCR(IPCR)was developed with Drosophila DNA under the condtions that only limit amount of DNA was available.The application of IPCR for analysis of unknown DNA sequences in individual flies show that flanking sequences can be amplified with only a half of genomic DNA from a single fly.This method may allow to clone unknown DNA sequences in single flies with the reliability and simplicity.