利用CODEHOP和iCODEHOP设计简并引物克隆芒果LAX基因家族片段

摘要

详细介绍采用CODEHOP和iCODEHOP设计简并引物的方法,并利用这2个程序设计了8对简并引物,从芒果子叶中扩增到8条不同长度的cDNA片段,测序结果表明,这些片段包含4类不同的DNA序列.经过在NCBI上进行blastx分析,这些序列与其它植物的LAX基因具有高度同源性,因此推测,芒果中的LAX基因家族其至少包括4条同源基因.研究结果表明CODEHOP和iCODEHOP设计的简并引物可信性强并具有高效性.%Rooting from mango cotyledon segment of zygote embryos was a good system to study the adventitious root formation mechanism. However, the molecular biology mechanism was unclear. In this study, the method of degenerate primers design using CODEHOP and iCODEHOP were detail introduced, and 8 pairs degenerate primes were selected for PCR. Then, 8 different cDNA fragments were amplified. Sequencing results showed that these fragments contained 4 different DNA sequences. After Blastx analysis on NCBI, it was found that these cDNAs belonged to one gene family and had high homology with other LAX genes reported. Therefore, there are at least 4 homogenous genes with LAX in mango. On the other hand, these results also suggested that degenerate primers designed by CODEHOP and iCODEHOP were of credibility and high efficiency.