CAJ | 학술논문

以‘甘肃红豆草’叶片总 RNA 为模板,通过 RT-PCR 方法扩增出肌动蛋白(Actin)基因片段,并克隆到pMD18-T载体,阳性克隆经PCR鉴定后测序.结果表明：序列分析表明,该片段长258 bp,编码86个氨基酸,所得序列与 GenBank中注册的Actin基因核苷酸序列的相似性在88%以上,与其他肌动蛋白的氨基酸序列的相似性达92%以上.注册该基因到 GenBank中,注册号为KP159623.采用半定量 RT-PCR方法,分析Actin基因在甘肃红豆草不同组织器官中的表达量相对恒定,而编码花青素还原酶的BAN基因在器官间的表达量差异较大,表明其可作为研究其它重要功能基因在‘甘肃红豆草’中的表达和调控的内参基因.
이‘감숙홍두초’협편총 RNA 위모판,통과 RT-PCR 방법확증출기동단백(Actin)기인편단,병극륭도pMD18-T재체,양성극륭경PCR감정후측서.결과표명：서렬분석표명,해편단장258 bp,편마86개안기산,소득서렬여 GenBank중주책적Actin기인핵감산서렬적상사성재88%이상,여기타기동단백적안기산서렬적상사성체92%이상.주책해기인도 GenBank중,주책호위KP159623.채용반정량 RT-PCR방법,분석Actin기인재감숙홍두초불동조직기관중적표체량상대항정,이편마화청소환원매적BAN기인재기관간적표체량차이교대,표명기가작위연구기타중요공능기인재‘감숙홍두초’중적표체화조공적내삼기인.
Total RNA of Onobrychis viciaefolia ‘Gansu’was extracted from leaf.The RNA was used to clone Actin gene fragment by RT-PCR,then the gene fragment was cloned into pMD1 8-T vector.The gene was sequenced after identifying the positives clone by PCR.The results revealed that the actin gene fragment from Onobrychis viciaefolia ‘Gansu’contained 258 bp and encoded a sequence of 86 amion acids.Similarity comparison showed that it shared over 88% similarity of nucleotide sequence similarity and over 92% similarity of amino acid sequence with other actin in the GenBank.The gene was submitted to GenBank and was given the accession number KP159623.Semi-quantitative PCR analysis indicated that the expression quantity of Actin gene was similar in different organs,however,the expression quantity of BAN gene encoding anthocyanin reductase was different.The results show that Actin gene can be used as the reference for analysis of gene expression in Onobrychis viciifolia ‘Gansu’.