Kondo R, Matsuura E T, Chigusa S I
Department of Biology, Ohcanomizu University, Tokyo, Japan.
Genet Res. 1992 Apr;59(2):81-4. doi: 10.1017/s0016672300030287.
By designing 3' ends of primers in PCR (polymerase chain reaction), a specific DNA fragment was selectively amplified in the presence of a 10(3)-fold excess of highly homologous (sequence difference ca. 2%) opponent DNA. This technique was applied in detecting paternal leakage of mitochondrial DNA (mtDNA) in intraspecific crosses of Drosophila simulans and interspecific crosses of Drosophila simulans and Drosophila mauritiana. The mtDNA types of their progeny were analysed by selective amplification of the paternal mtDNA fragment possessing a polymorphic restriction site and detecting its cleaved fragments. Paternal mtDNA was detected in the progeny of 14 out of 16 crosses. The present result indicates small but frequent inheritance of sperm mtDNA in Drosophila, which is supportive to our previous finding.
通过在聚合酶链反应(PCR)中设计引物的3'端,在存在10³倍过量的高度同源(序列差异约2%)的对立DNA的情况下,选择性地扩增了特定的DNA片段。该技术被应用于检测拟暗果蝇种内杂交以及拟暗果蝇与毛里求斯果蝇种间杂交中线粒体DNA(mtDNA)的父系渗漏。通过对具有多态性限制性位点的父系mtDNA片段进行选择性扩增并检测其切割片段,分析了它们后代的mtDNA类型。在16个杂交组合中的14个后代中检测到了父系mtDNA。目前的结果表明,果蝇中精子mtDNA的遗传虽少但频繁,这支持了我们之前的发现。