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根据丝氨酸/苏氨酸蛋白激酶类催化结构域Ⅰ和Ⅸ的氨基酸保守序列设计简并引物,PCR扩增‘紫花芒’杧果嫩绿色叶片基因组DNA。在NCBI中用BlastX比对发现,有6个阳性克隆是抗病基因同源序列,并命名为PK1~PK6,GenBank注册序列号为AY693369~AY693371和AY776277~AY776279。在PK3、PK4和PK6中发现了丝氨酸/苏氨酸蛋白激酶的9个催化结构域Ⅰ~Ⅸ,而其它3个克隆具有部分结构域。氨基酸序列同源性分析还发现,6个克隆编码的氨基酸序列都与受体样蛋白激酶有较高的氨基酸序列同源性。系统进化树分析还表明它们都是可能的抗病基因同源序列。总之,6个克隆不仅是抗病基因同源序列而且是可能的受体样蛋白激酶基因同源序列。
Degenerate primers were designed based on the conserved amino acid sequences of serine / threonine protein kinase catalytic domains Ⅰ and Ⅸ, and genomic DNA of tender green leaves of ’Vetch’ was PCR amplified. In the NCBI BlastX comparison found that six positive clones are disease-resistant gene homologous sequences, and named PK1 ~ PK6, GenBank accession numbers AY693369 ~ AY693371 and AY776277 ~ AY776279. Nine catalytic domains of serine / threonine protein kinases I to IX were found in PK3, PK4 and PK6, while the other three had partial domains. Amino acid sequence homology analysis also found that all six amino acid sequences encoded by the receptor-like protein kinase have high amino acid sequence homology. Phylogenetic tree analysis also showed that they are all possible resistance gene homologous sequences. In summary, the six clones are not only homologous sequences of resistance genes but also possible receptor-like protein kinase gene homologous sequences.