植物高度同源重复基因的PCR检测

Detection of highly homologous genes in plants with PCR

以油菜中三个同源性高达90%以上的AP3基因为研究对象,采用三种不同的DNA聚合酶进行常规PCR扩增和纳米PCR扩增.结果显示:纳米PCR比普通Taq DNA聚合酶和低保真Plus DNA聚合酶具有更高的扩增特异性,即在相同的退火温度下,只有纳米粒子PCR扩增出特异性的目的产物而无非特异性产物;虽然高保真Pfu DNA聚合酶PCR显示出了同纳米PCR一样的扩增特异性,但纳米PCR不仅能表现出较高的扩增特异性,而且还显示出了更好的扩增效率.并且,纳米PCR即使是在30℃的低退火温度下依旧能扩增出特异性的目的条带,表现出纳米PCR良好的扩增特异性和扩增效率.这些结果表明,纳米PCR在基因检测等方面具有一定的优势.

Three homologous AP3 genes with more than 90% identity in Brassica napus as the research object, three different DNA Polymerases and nanoparticles were used in the PCR reaction to detect the efficiency of plant duplicate gene amplification. The results showed that the nano-particles have a higher PCR amplification specificity compared with the common TaqDNA polymerase and low-fidelity PlusDNA polymerase. Even at the same annealing temperature, only nano-particles PCR amplified spe- cific products without non-specific products amplification. Although high-fidelity Pfu polymerase PCR showed the same PCR amplification specificity with Nano-particles, Yet Nano-particles PCR show a higher PCR amplification specificity and good amplification efficiency. And nano-particles even at low annealing temperature of 30℃ is still able to amplify specific purpose of bands, demonstrated its good amplification specificity and efficiency. These results indicate that nano-particles PCR for gene detection has certain advantages.