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两个烟草par A相关基因在转基因植物中的器官特异性和生长素诱导型表达。

Organ-specific and auxin-inducible expression of two tobacco par A-related genes in transgenic plants.

作者信息

Niwa Y, Muranaka T, Baba A, Machida Y

机构信息

Department of Biology, Faculty of Science, Nagoya University, Japan.

出版信息

DNA Res. 1994;1(5):213-21. doi: 10.1093/dnares/1.5.213.

Abstract

We have isolated four genomic DNA clones that contain the transcription initiation site of the parA gene(s) from a tobacco genomic library by hybridization with the 5' segment of the parA cDNA previously isolated. They were classified into two types on the basis of their nucleotide sequences. Southern blot analysis indicated that two types of clones were respectively derived from the two parental species of tobacco, Nicotiana tomentosiformis and Nicotiana sylvestris. The genes corresponding to these clones were designated as parAt and parAs, respectively, and the parA cDNA clone was shown to code for mRNA from parAt on the basis of its nucleotide sequence. The 5' regions about 400 nucleotides upstream from the transcription initiation sites of the parAt and parAs genomic clones were highly homologous to one another, but regions further upstream showed no significant similarity. The coding sequence of the GUS (beta-glucuronidase) reporter gene was linked to the 5'-upstream regions of parAt and parAs, and the sites of expression of these fusion genes were examined in transgenic tobacco plants. In the absence of auxins, both fusion genes were expressed in capsules at a late stage of seed development, mature seeds, a root apex and a root-hair zone whereas no significant expression was seen in other organs. Their expression was enhanced by 2,4-dichlorophenoxyacetic acid in most of the organs of tobacco. The results show that expression of these genes is regulated by both organ-specific and auxin-inducible mechanisms.

摘要

我们通过与先前分离的parA cDNA的5'片段杂交,从烟草基因组文库中分离出四个基因组DNA克隆,这些克隆包含parA基因的转录起始位点。根据它们的核苷酸序列,将它们分为两种类型。Southern印迹分析表明,这两种类型的克隆分别来自烟草的两个亲本物种,即绒毛状烟草(Nicotiana tomentosiformis)和林烟草(Nicotiana sylvestris)。与这些克隆相对应的基因分别被命名为parAt和parAs,并且根据其核苷酸序列表明parA cDNA克隆编码来自parAt的mRNA。parAt和parAs基因组克隆的转录起始位点上游约400个核苷酸的5'区域彼此高度同源,但更上游的区域没有明显的相似性。将GUS(β-葡萄糖醛酸酶)报告基因的编码序列连接到parAt和parAs的5'上游区域,并在转基因烟草植物中检测这些融合基因的表达位点。在没有生长素的情况下,两个融合基因在种子发育后期的荚果、成熟种子、根尖和根毛区中表达,而在其他器官中未观察到明显的表达。在烟草的大多数器官中,它们的表达被2,4-二氯苯氧乙酸增强。结果表明,这些基因的表达受器官特异性和生长素诱导机制的调节。

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