克隆植物苯丙氨酸解氨酶基因通用引物的发掘与应用

根据松树、咖啡、山茶、鳄梨和白杨PAL基因第2外显子的保守区段,设计了1对简并引物。以该引物对88种植物的基因组DNA进行了PCR扩增。结果发现,每种植物中都能扩增出介于750～1 000 bp的特异片段。随机选取银杏、夹竹桃、核桃、海桐和盘槐的PCR扩增产物进行测序分析,发现其大小分别为862 bp、866 bp、866 bp、866 bp和865 bp。使用生物信息学进行进化树分析,所克隆的5种植物的特异片段与其亲缘关系密切的物种PAL高度同源,初步证实了这些片段是银杏、夹竹桃、核桃、海桐和盘槐的PAL基因。因此,本研究开发的PAL基因简并引物,可以克隆任意植物的PAL基因。

Excavation and Application of Universal Primers for Phenylalanine Ammonia-lyase Gene Cloning from Plants

A pair of degenerate primers were disigned based on the conserved sequences of exon II of phenylalanine ammonia-lyase（PAL）gene of pinus,coffee,camellia,avocado and white poplar,which were used in PCR amplification of genomic DNA extracted from 88 kinds of plants,respectively.The results showed that the specific fragments between 750 bp to 1 000 bp were obtained from all of the plants.Five PCR products were randomly selected to DNA sequencing,and the fragments were 862 bp,866 bp,866 bp,866 bp and 865 bp,respectively.The fragments were highly homologous to PAL gene of plants closely related to them,which suggested that they were all PAL.Therefore,the degenerate primers specific could used in PAL gene cloning from all plants.