CAJ | 학술논문

目的：克隆刺五加Actin基因的全长cDNA序列，并对其进行生物信息学分析。方法：以刺五加的叶片为材料，提取总RNA，逆转录为cDNA，根据初步克隆到的刺五加Actin基因保守序列设计引物，利用套式PCR进行3'和5'RACE扩增，得到Actin基因的3'和5'端cDNA序列片段，拼接获得cDNA全长。将该序列进行BLAST比对、相似度分析，并对刺五加Actin1蛋白的二级结构和三级结构进行预测。结果：刺五加的Actin基因全长1507 bp，命名为ESA ctin1，注册号KC469585。该基因开放阅读框（ORF）长1134 bp，编码377个氨基酸，5'端非编码区长140 bp，3'端非编码区长233 bp。ESActin1与GenBank 中注册的其它植物肌动蛋白核苷酸序列的相似性在75%以上，蛋白质序列的相似性在94%以上。结论：首次报道了刺五加Actin基因的全长cDNA序列，为刺五加的分子生物学研究奠定基础。
목적：극륭자오가Actin기인적전장cDNA서렬，병대기진행생물신식학분석。방법：이자오가적협편위재료，제취총RNA，역전록위cDNA，근거초보극륭도적자오가Actin기인보수서렬설계인물，이용투식PCR진행3'화5'RACE확증，득도Actin기인적3'화5'단cDNA서렬편단，병접획득cDNA전장。장해서렬진행BLAST비대、상사도분석，병대자오가Actin1단백적이급결구화삼급결구진행예측。결과：자오가적Actin기인전장1507 bp，명명위ESA ctin1，주책호KC469585。해기인개방열독광（ORF）장1134 bp，편마377개안기산，5'단비편마구장140 bp，3'단비편마구장233 bp。ESActin1여GenBank 중주책적기타식물기동단백핵감산서렬적상사성재75%이상，단백질서렬적상사성재94%이상。결론：수차보도료자오가Actin기인적전장cDNA서렬，위자오가적분자생물학연구전정기출。
This study was aimed to clone the full-length cDNA sequence of actin gene of Eleutherococcus sentico-sus. And bioinformatics analysis was used. The total RNA was isolated from leaves of E. senticosus , and cDNA was synthesized by reverse transcription of total RNA. Primers were designed according to the conserved se-quence that had been cloned of Actin of E. senticosus . Then, the 3'and 5' cDNA fragments were cloned by nested PCR . The full-length gene was obtained by gene splicing method . Sequencing results were compared and treated with similarity analysis by blast analysis in the GenBank. Protein secondary structure and tertiary struc-ture of Actin of E. senticosus was predicted by online software. The results showed that the full-length cDNA of Actin of E. senticosus is 1507 bp, which named EsActin1, GenBank accession No. KC469585. The conserved sequence, which contained a 1134 bp open reading frame that encoding a 377 amino acid residues, a 5'-UTR of 140 bp and a 3'-UTR of 233 bp. Homologous alignment showed that it shared over 75% nucleotide se-quence similarity and over 94% amino acid sequences similarity with Actins in other plants. It was concluded that this study first isolated and reported the full-length cDNA sequence of actin gene of E. senticosus , and laid a foundation for the molecular biology research of E. senticosus .