Guan Cuiping, Zhou Xueping
State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310029, PR China.
Virus Res. 2006 Feb;115(2):150-7. doi: 10.1016/j.virusres.2005.08.002. Epub 2005 Sep 19.
DNAbeta is a satellite molecule associated with some monopartite begomoviruses and encodes a single gene (betaC1), which is highly conserved in position and size among DNAbeta molecules. A 955 nt fragment of Tomato yellow leaf curl China virus (TYLCCNV) DNAbeta, upstream of the translation start site of betaC1 gene was tested for its promoter activity with gus as a reporter gene. Analysis of beta-glucuronidase (GUS) activity following transient expression assays indicated that the 955 nt fragment had promoter activity and that 3'-deletions of 399 or 173 nt of the fragment resulted in complete loss of its promoter activity. The 5'-deletions of 782 or 556 nt of the fragment, however, did not affect its activity. Histochemical staining revealed that this fragment can be used to express gus gene specifically in phloem tissue of stably transformed tobacco plants. Further studies have indicated that a 173 nt segment from 3'-end of the 955 nt fragment was responsible for basic promoter activity and phloem-specific expression.
DNAβ是一种与某些单分体双生病毒相关的卫星分子,它编码一个单一基因(βC1),该基因在DNAβ分子中的位置和大小高度保守。以gus作为报告基因,对番茄黄化曲叶中国病毒(TYLCCNV)DNAβ中βC1基因翻译起始位点上游的一段955 nt片段进行了启动子活性检测。瞬时表达分析后对β-葡萄糖醛酸酶(GUS)活性的分析表明,该955 nt片段具有启动子活性,并且该片段399 nt或173 nt的3'-缺失导致其启动子活性完全丧失。然而,该片段782 nt或556 nt的5'-缺失并不影响其活性。组织化学染色显示,该片段可用于在稳定转化的烟草植株的韧皮部组织中特异性表达gus基因。进一步研究表明,955 nt片段3'-末端的一段173 nt序列负责基本启动子活性和韧皮部特异性表达。