豌豆Le启动子的功能的研究_百替生物.pdfVIP

Le Le 豌豆 启动子的功能研究 LLee ProductionofthePromoterofLe(Lengthgene)inGarden ProductionofthePromoterofLe(Lengthgene)inGarden PPrroodduuccttiioonnoofftthheePPrroommootteerrooffLLee((LLeennggtthhggeennee))iinnGGaarrddeenn Pea Pea PPeeaa 生命科学学院97级 王萌 摘要 用PCR的方法从豌豆总DNA中克隆了Le基因（lengthgene）的启动子，命名为Le 启动子。Northern 杂交的结果证明，Le基因主要在豌豆的根、茎中表达，在花和叶中的 表达量很少，而果实中几乎没有表达。根据Le 基因伸长组织特异的表达图式，推测Le 基因的启动子为伸长组织特异的启动子。GFP基因作为报告基因，由Le启动子调控，在 转基因烟草中表达。对转基因烟草叶片的荧光显微镜观察结果，以及用GFP基因作为探 针，转基因烟草的Northern 杂交的结果，证实了Le 启动子的组织特异性。利用Le 启动 子，可以实现目的基因的组织特异表达，在理论研究和生产实践上都有重要的应用价值。 Abstract Stem length is one of the seven traits of garden pea from which Gregor Mendel established the laws of inheritance. The length gene (Le) encodes 3β-hydroxylase, which controls the GA1 biosynthesis from GA in pea. GA is the major gibberellin (GA) controlling stem elongation 20 1 in pea (PisumsativumL.).The Northern Blotting analysis showed the tissue-specific expression of Legene in roots and shoots, using the coding sequence of Legene as a probe. Based on the service@100 service@100 wwwwww..110000bbiiootteecchh..ccoomm sseerrvviiccee@@110000bbiiootteecchh..ccoomm result, the promoter of Legene shoμLd have tissue-specificity. Using PCR (Polymerase Chain Reaction), we cloned the promoter of Legene (Lepromoter) from the genomic DNAof G2 pea. Under the control of the Lepromoter, the GFPgene was expressed in tobacco. Using the GFP gene as a probe, the Northern Blotting analysis of transgenic tobacco showed that the GFPgene expressed specifically in roots and