GUS基因和NPTII基因表达的相关性及其在转基因棉花检测研究中的应用  

Expression of GUS Gene and NPTII Gene and Their Application in the Detecting of Transgenic Cotton

作　　者：上官小霞[1] 李燕娥[1] 梁运生[1] 吴霞[1] 杜春芳[1] 张林水[1] 

机构地区：[1]山西省农业科学院棉花研究所,山西运城044000

出　　处：《棉花学报》2007年第3期163-167,共5页Cotton Science

基　　金：国家"863"项目(项目编号2004AA212102)

摘　　要：利用农杆菌介导转化法,将含有35S启动子驱动NPTII基因和GUS基因以及棉纤维特异表达启动子E6驱动目的基因FB的植物表达载体转入到常规棉花R15中。重点分析了GUS基因和NPTII基因在愈伤诱导阶段、T0代及T1代转基因棉花中的表达情况。综合两个基因的表达来进行转基因棉花的阳性鉴定,可以为转基因棉花后代的纯合选育提供双重保障。7个转基因株系选育到T3代共获得株行51个,卡那霉素检测多数株行阳性率在90%以上,其中21个株行阳性率达100%。By Agrobacterium-mediated method, the plant expression vector contained the target gene FB driven by a cotton fiber specific expression promoter E6 was transferred into cotton cultivar R15. The vector also contains a GUS gene,and a NPTII gene driven by 35S promoter, respectively. The expression of GUS gene and NPTII gene was analyzed in the stage of resistant calli selected, To and T1 generation transgenic plants. These two genes are in the same T-DNA in the vector, their expression has high relativity. Because the phenomena of transgenic silencing and the effect of insert sites, the expression of GUS gene and NPTII gene has some differences. We obtained the resistant calli through detecting the expression of GUS gene and NPTII gene together. Kanamycin resistance, GUS staining, and PCR detection were conducted on the 15 putative To generation transgenic plants. Among them, 11 plants were positive for the transgene according To PCR detection, the positive ratio reached 73.3 %. 12 plants showed kanamycin resistance, 10 plants were GUS-staining positive, the expression of NPTII gene and GUS gene showed high relativity. 10 positive plants were obtained through the three derected methods among the 15 To generation transgenic plants. The breeding of transgenic progeny also relied on both kanamycin examination and GUS staining. Of T1 transgenic progeny, the expression of GUS gene and NPTII gene has significant difference, so there has some limitation for To as a certain extent for screening the transgenic progeny only by kanamycin examination. Kanamycin assay was performed in the field, climate and other circumstance -factors may influence the result. However, being simple and fast, this assay can employed as a preliminary screen, three times of kanamycin assay were performed during the period of cotton vegetable development, and then GUS staining were acted based on the result of kanamycin resistant test. Proceeded the two methods together , the limitation of each measure can be compensated, accordingly the efficienc

关 键 词：GUS基因 NPTII基因 相关性 转基因棉花 基因表达 

分 类 号：S562[农业科学—作物学]