香蕉束顶病毒(BBTV)基因组至少由6个大小约为1.0-1.1kb的单链环状DNA组分所组成,每一个DNA组分包含编码区与非编码区。本文在前人的研究基础上进一步了解BBTV DNA组分启动子的功能。首先根据BBTV 海南分离物的全序列,通过常规PCR扩增出长为540bp的 BBTV DNA3组分启动子序列BV3.1,同时通过重叠PCR扩增出646bp的DNA2与DNA3组分非编码区拼接的重组启动子序列BV23,分别替代pBI121 35S启动子序列与gus基因进行融合,构建植物表达载体pBIBV3.1、pBIBV23。农杆菌介导转化获得的pBIBV3.1转基因烟草经GUS化学组织染色后,在其叶片的叶脉处检测到微弱的GUS活性,证实了DNA3组分的韧皮部特异表达活性;而pBIBV23转基因烟草,其叶片经GUS组织化学染色后,在叶肉、叶缘及一些叶脉上检测到弱GUS活性,这表明由BV23驱动的gus基因在烟草中类似于组成型表达,则DNA2组分转录方式可能有异于DNA3组分。
Banana bunchy top disease is one of major diseases in banana, the pathogen of the disease is banana bunchy top virus. The virus genome consists of at least 6 components of circular ssDNA about 1.0~1.1kb. Each DNA component contains encoding region and non-coding one. On the basis of previous research, we want to understand further the promoter function of BBTV DNA components. According as the genomic sequence of BBTV Hainan isolate, a 540bp fragment(BV3.1) generated by PCR amplification, which is the promoter sequence of BBTV DNA3 component; and a 646bp splicing sequence(BV23)is consisted of non-coding regions of DNA2 and DNA3 components generated by overlapping PCR amplification. This two promoter sequences substituted the 35S promoter sequence in pBI121 vector and fuse with gus gene respectively, and then got the plant expression vector named for pBIBV3.1(540bp)、pBIVBV23(646bp) are constructed. By GUS histochemical staining, the very weak GUS activity was limited in the leave veins of pBIBV3.1 transgenic tobacco, it confirmed the promoter activity is associated with phloem specific expression. Whereas the faint GUS activities were detected in the mesophyll, the leaf margin of pBIVBV23 transgenic tobacco and in some leaf veins as well. It indicates that the transcriptional method of DNA2 component is different with one of DNA3 component.
庄军,刘志昕. 香蕉束顶病毒DNA组分2、3的启动子区的组织特异性分析[J]. 中国生物工程杂志, 2007, 27(9): 69-73.
. Analysis of the promoter derived tissue specific activity about banana bunchy top virus DNA2 and DNA3 components. China Biotechnology, 2007, 27(9): 69-73.
https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2007/V27/I9/69
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