Bhattacharyya-Pakrasi M, Peng J, Elmer J S, Laco G, Shen P, Kaniewska M B, Kononowicz H, Wen F, Hodges T K, Beachy R N
Department of Biology, Washington University, St. Louis, MO 63130.
Plant J. 1993 Jul;4(1):71-9. doi: 10.1046/j.1365-313x.1993.04010071.x.
The major promoter region for the transcription of the genome of rice tungro bacilliform virus (RTBV), a newly described badnavirus, has been identified. Fragments of the RTBV genome upstream of the site of transcription initiation were isolated and tested for promoter activity using a beta-glucuronidase receptor gene (gusA). Assays of transient gusA expression were performed following introduction of the chimeric gene into protoplasts via electroporation. The chimeric RTBV-promoter: gusA gene was more active in rice protoplasts than in maize or tobacco protoplasts, but was weaker than gusA controlled by an enhanced 35S promoter from cauliflower mosaic virus. Analysis of gusA gene expression following introduction of chimeric reporter genes into intact leaves via micro-projectile bombardment indicated that the GUS activity is present primarily in vascular tissues. Transgenic rice plants carrying the chimeric gusA gene had GUS activity only in the phloem of the vascular bundles in the leaf. Tissue printing studies demonstrated that RTBV accumulates in the vascular bundles of infected rice leaves. The results of our study indicate that phloem-specific expression from the RTBV promoter is an intrinsic property of the viral promoter.
一种新发现的杆状DNA病毒——水稻东格鲁杆状病毒(RTBV)基因组转录的主要启动子区域已被确定。分离了RTBV基因组转录起始位点上游的片段,并使用β-葡萄糖醛酸酶报告基因(gusA)检测其启动子活性。通过电穿孔将嵌合基因导入原生质体后,进行瞬时gusA表达分析。RTBV启动子:gusA嵌合基因在水稻原生质体中的活性高于玉米或烟草原生质体,但弱于由花椰菜花叶病毒增强型35S启动子控制的gusA。通过微粒轰击将嵌合报告基因导入完整叶片后对gusA基因表达的分析表明,GUS活性主要存在于维管组织中。携带嵌合gusA基因的转基因水稻植株的GUS活性仅存在于叶片维管束的韧皮部中。组织印迹研究表明,RTBV在受感染水稻叶片的维管束中积累。我们的研究结果表明,RTBV启动子的韧皮部特异性表达是病毒启动子的固有特性。