西北植物学报
西北植物學報
서북식물학보
ACTA BOTANICA BOREALI-OCCIDENTALIA SINICA
2010年
1期
1-7
,共7页
二氢黄酮还原酶%植物表达载体构建%转化
二氫黃酮還原酶%植物錶達載體構建%轉化
이경황동환원매%식물표체재체구건%전화
dihydroflavonol reductase gene%construction of plant expression vector%transformation
以蒺藜苜蓿(Medicago trunctula cv.5160)幼果总RNA为模板,采用RT-PCR技术克隆到二氢黄酮还原酶(DFR)基因的cDNA序列,所获得的cDNA序列全长1 018 bp,具有完整的ORF,编码337个氨基酸.Blast分析表明,该片段与GenBank中注册的DFR基因同源性为99.80%.以植物表达载体pBI121为基础,构建了CaMV 35S启动子驱动的DFR基因植物表达载体pBIDFR;采用直接转化法将pBIDFR导入根癌农杆菌EHA105,用该菌株对普通烟草进行遗传转化获得6株转基因植株.
以蒺藜苜蓿(Medicago trunctula cv.5160)幼果總RNA為模闆,採用RT-PCR技術剋隆到二氫黃酮還原酶(DFR)基因的cDNA序列,所穫得的cDNA序列全長1 018 bp,具有完整的ORF,編碼337箇氨基痠.Blast分析錶明,該片段與GenBank中註冊的DFR基因同源性為99.80%.以植物錶達載體pBI121為基礎,構建瞭CaMV 35S啟動子驅動的DFR基因植物錶達載體pBIDFR;採用直接轉化法將pBIDFR導入根癌農桿菌EHA105,用該菌株對普通煙草進行遺傳轉化穫得6株轉基因植株.
이질려목숙(Medicago trunctula cv.5160)유과총RNA위모판,채용RT-PCR기술극륭도이경황동환원매(DFR)기인적cDNA서렬,소획득적cDNA서렬전장1 018 bp,구유완정적ORF,편마337개안기산.Blast분석표명,해편단여GenBank중주책적DFR기인동원성위99.80%.이식물표체재체pBI121위기출,구건료CaMV 35S계동자구동적DFR기인식물표체재체pBIDFR;채용직접전화법장pBIDFR도입근암농간균EHA105,용해균주대보통연초진행유전전화획득6주전기인식주.
The dihydroflavonol reductase (DFR) gene was cloned from young fruit of Medicago truncatula cv.5160 by RT-PCR and the coding sequences of this gene were analyzed.The result indicated that the full-length cDNA is 1 018 bp and contain open reading frame (ORF) encoding 337 amino-acids.The sequences were analyzed with software Blast and exhibited homologous 99.80% with DFR from GenBank.Then the promoter CaMV35S driven,plant expression vector pBIDFR with DFR was constructed based on the vector pBI121.By direct DNA transfer,pBIDFR was transferred into Agrobacterium tumefacien EHA105.Then transfer the new engineering bacterium to Nicotiana tabacum and six transgenic tobacco plants were obtained.
