CAJ | 학술논문

根据NCBI网站上萼脊兰AP1-like全长基因序列设计引物,采用RT-PCR法从萼脊兰花瓣中扩增SeAP1-like基因,对扩增产物进行克隆和测序。结果表明,获得的萼脊兰SeAP1-like基因为743 bp,编码247个氨基酸。将SeAP1-like基因插入pCAMBIA1304载体中,经PCR、双酶切鉴定,证实重组表达质粒中含有目的片段,表明成功构建了高效植物表达载体 pCAMBIA1304-SeAP1。对SeAP1-like基因的结构域和所表达蛋白质的亲水性进行了分析,并预测了该基因所表达蛋白的二维结构。结果显示,该基因包含MADS-box和K-box保守域,属于MADS-box基因家族,该蛋白分子属于亲水性蛋白。二级结构分析表明,其包含57.49%的α螺旋、6.07%的延伸链、36.44%的不规则折叠。
근거NCBI망참상악척란AP1-like전장기인서렬설계인물,채용RT-PCR법종악척란화판중확증SeAP1-like기인,대확증산물진행극륭화측서。결과표명,획득적악척란SeAP1-like기인위743 bp,편마247개안기산。장SeAP1-like기인삽입pCAMBIA1304재체중,경PCR、쌍매절감정,증실중조표체질립중함유목적편단,표명성공구건료고효식물표체재체 pCAMBIA1304-SeAP1。대SeAP1-like기인적결구역화소표체단백질적친수성진행료분석,병예측료해기인소표체단백적이유결구。결과현시,해기인포함MADS-box화K-box보수역,속우MADS-box기인가족,해단백분자속우친수성단백。이급결구분석표명,기포함57.49%적α라선、6.07%적연신련、36.44%적불규칙절첩。
The primers were designed according to the reference Sedirea japonica AP1-like gene from NC-BI. The SeAP1-like gene was amplified from the scape of Sedirea japonica petal by RT-PCR. The PCR product was sequenced. The sequencing result showed the target gene was 743 bp encoding 247 putative amino acid residues. The recombinant plasmid pCAMBIA1304-SeAP1 was obtained by identified by inser-ting the SeAP1-like gene into pCAMBIA1304 vector,and identified by PCR,double digestion. The results showed that the recombinant plasmid pCAMBIA1304-SeAP1 which contained the target gene was con-structed successfully. The structural domain of the SeAP1-like gene and the hydrophily of the protein ex-pressed by the SeAP1-like gene were analyzed,and the secondary structure of the protein expressed by the SeAP1-like gene was predicted. The results showed that SeAP1-like gene contained the MADS-box and K-box domain,belonged to MADS-box super family. This protein expressed by the SeAP1-like gene was hydro-philic and contained 57. 49% α-helical domains,6. 07% extended strand,36. 44% random coil.