CAJ | 학술논문

【目的】克隆蝴蝶兰MADS-Box基因并构建其正义和反义植物表达载体，为其功能研究奠定基础。【方法】以蝴蝶兰杂交品种（Phalaenopsis hybrid cv.Jiuhbao Red Rose）为试验材料，采用RT-PCR和RACE技术从花葶中克隆MADS-Box基因，并构建正义和反义植物表达载体。【结果】克隆获得一个蝴蝶兰MADS-Box基因，命名为DtpsMADS1（GeneBank登录号JQ065097）。该基因cDNA全长960 bp，包含37 bp的5'非编码区、185 bp的3'非编码区和一个738 bp的编码区；该基因编码245个氨基酸。生物信息学分析结果表明，该基因编码的蛋白质为碱性亲水性蛋白，具有62.45%的α-螺旋，8.16%的延伸链和29.39%的不规则折叠。序列比对和系统进化分析结果表明，DtpsMADS1与蝴蝶兰ORAP13的亲缘关系最近，同源性达99.0%，与石斛和蕙兰的同源性分别为83.0%和82.0%，属于MADS家族A类亚家族。将DtpsMADS1基因连接到植物表达载体pBI121上，构建获得正、反义植物表达载体pBI121-DtpsMADS1-S和pBI121-DtpsMADS1-A。【结论】成功克隆的蝴蝶兰DtpsMADS1基因属于MADS家族A类亚家族，具有明显的保守性和特异性，可为蝴蝶兰DtpsMADS1基因功能的鉴定及蝴蝶兰的遗传改良奠定基础。
【목적】극륭호접란MADS-Box기인병구건기정의화반의식물표체재체，위기공능연구전정기출。【방법】이호접란잡교품충（Phalaenopsis hybrid cv.Jiuhbao Red Rose）위시험재료，채용RT-PCR화RACE기술종화정중극륭MADS-Box기인，병구건정의화반의식물표체재체。【결과】극륭획득일개호접란MADS-Box기인，명명위DtpsMADS1（GeneBank등록호JQ065097）。해기인cDNA전장960 bp，포함37 bp적5'비편마구、185 bp적3'비편마구화일개738 bp적편마구；해기인편마245개안기산。생물신식학분석결과표명，해기인편마적단백질위감성친수성단백，구유62.45%적α-라선，8.16%적연신련화29.39%적불규칙절첩。서렬비대화계통진화분석결과표명，DtpsMADS1여호접란ORAP13적친연관계최근，동원성체99.0%，여석곡화혜란적동원성분별위83.0%화82.0%，속우MADS가족A류아가족。장DtpsMADS1기인련접도식물표체재체pBI121상，구건획득정、반의식물표체재체pBI121-DtpsMADS1-S화pBI121-DtpsMADS1-A。【결론】성공극륭적호접란DtpsMADS1기인속우MADS가족A류아가족，구유명현적보수성화특이성，가위호접란DtpsMADS1기인공능적감정급호접란적유전개량전정기출。
[Objective]Cloning as well as sense and antisense plant expression vector of MADS-Box gene in Phalaenopsis hybrid were conducted to provide references for researching its functions. [Method]The MADS-Box gene was cloned from scape of Phalaenopsis hybrid cv. Jiuhbao Red Rose using RT-PCR and RACE. Sense and antisense plant expression vector were also constructed. [Result]The cloned MADS-Box gene from scape was named DtpsMADS1(GenBank accession No. JQ065097). The full length cDNA of DtpsMADS1 was 960 bp, containing 37 bp of a 5'-untranslated region (5'-UTR),185 bp of 3'-UTR,and 738 bp of an opening reading frame (ORF). The gene encoded 245 predicted amino acids. The bioinfor-mation analysis results showed that the gene encoding protein was a basic hydrophilic protein and contained 62.45%ofα-heli-cal domains, 8.16%of extended strand and 29.39%of random coil. Sequence comparison and phylogenetic analysis revealed that DtpsMADS1 shared 99.0%homology with ORAP13 of Phalaenopsis amabilis, 83.0%and 82.0%homology with Dendrobi-um nobile and Cymbidium orchid, respectively. DtpsMADS1 belonged to A-class subfamily of MADS family. Sense and anti-sense plant expression vector pBI121-DtpsMADS1-S and pBI121-DtpsMADS1-A were constructed when DtpsMADS1 was connected to plant expression vector pBI121. [Conclusion]The DtpsMADS1 gene of Phalaenopsis hybrid was cloned suc-cessfully. The sense and antisense plant expression vector were constructed successfully and could be used for DtpsMADS1 genetic function identification and genetic improvement of Phalaenopsis hybrid.