燕麦Actin基因片段的克隆及序列分析

摘要

Obj ective]To clone and analyze the Actin gene from Avena sativa.[Method]Degenerate primers were designed based on the conserved sequences of the actin genes from other plants.Total RNA was extracted from the leaves of A.sativa.Actin gene fragment was obtained by reverse transcription poly-merase chain reaction (RT-PCR)and cloned into PUCm-T vector.The positive clone identified by PCR was sequenced.[Result]The amplified fragment from A.sativa contains 678 bp,encoded a protein of 225 amino acids.Homology comparison with other plants actin gene sequences in the GenBank showed that it shared over 85% nucleotide sequence homology and 93%amino acid sequence homology with other plants actins.[Conclusion]Actin gene of A.sativa has an intimate genetic relationship with Actin genes of Leymus chinensis,Lophopyrumelongatum and Loliumperenne.%【目的】克隆与分析燕麦肌动蛋白基因片段．【方法】根据其他植物Actin基因的保守序列设计一对简并性引物，以燕麦叶片总RNA为模板，采用RT-PCR的方法扩增出Actin基因片段并克隆到PUCm-T载体，进而转化大肠杆菌E．coli DH5α感受态细胞．阳性克隆经PCR鉴定后进行测序．【结果】扩增片段长678 bp，共编码225个氨基酸；所得序列与GenBank中注册的Actin基因序列的同源性均在85％以上，与其他肌动蛋白的氨基酸序列的同源性达93％以上．【结论】系统进化分析表明，扩增到的燕麦肌动蛋白基因与羊草、长穗偃麦草和黑麦草等植物的肌动蛋白基因亲缘关系最为密切．