摘要
内含子是基因的重要组成部分,它与功能基因表达之间的关系越来越被重视。本研究以pCAM-BIA3301为载体,利用玉米泛素启动子Ubi1和水稻肌动蛋白启动子Actin1构建两个GUS基因表达载体p33U1和p33A1,同时设置3个对照载体。通过基因枪轰击法将上述载体转入水稻胚性愈伤组织,探讨内含子对外源目的基因表达的调控作用。组织化学检测结果表明:CaMV35S启动子调控下的iGUS(带内含子)基因能够顺利表达;同样,Actin1启动子(带内含子)调控下的不带内含子的GUS基因也可以正常表达,而当Actin1启动子(带内含子)驱动iGUS基因时,则导致GUS染色反应不能发生。Ubi1启动子(带内含子)调控GUS基因的瞬时表达也得出类似结果,证明表达框中内含子的数量为1个或两个时,对GUS基因的表达起到了不同的调控作用。本研究结果对植物表达载体构建及功能基因表达都具有指导意义。
As one of important component of plant genome,intron is attracting more and more attention in the relation with functional gene expression.In the present work,two vectors p33U1 and p33A1 which harboring GUS genes were constructed based on the primary vector pCAMBIA3301,together with other three vectors as the controls,were introduced into rice calli via particle bombardment to investigate the GUS expression pattern affected by numbers of intron in the expression cassette.The results of histochemical assay showed that when GUS gene with an intron inside was driven by CaMV35S promoter,GUS protein can be expressed normally and detected by staining.Similar result can also be obtained when the GUS gene without intron inside was driven by Actin1 promoter bearing an intron.However,when the iGUS gene with an intron was regulated by Actin1 promoter,resulting in the existence of two introns closely in the expression cassette,GUS protein will not be detectable.Another Ubi promoter having an intron demonstrated the similar result.Evidences in this study prove that one or two introns in expression cassette resulted different transient expression patterns of GUS gene,and guide researchers in vector construction and gene expression of plants.
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2011年第5期571-576,共6页
Genomics and Applied Biology
基金
吉林省科技发展重大项目(20065008)
吉林省博士后基金共同资助
