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Floral organ specific expression promoter separated from Populus trichocarpa Torr.&Gray and application thereof

A flower organ and promoter technology, applied in the field of promoters, can solve the problems of hidden safety concerns of transgenic plants, unfavorable plant growth, excessive consumption of cell materials and energy, etc.

Active Publication Date: 2014-11-05
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the promoters used in plant genetic engineering are constitutive promoters, which enable high-level expression of exogenous target genes in various plant tissues. Space effectively regulates the expression of target genes, excessive consumption of intracellular material and energy (Gittins JR, Pellny TK, Hiles ER, Rosa C, Biricolti S, et al. (2000) Transgene expression driven by heterologous ribulose-1,5 -bisphosphate carboxylase / oxygenase small-subunit gene promoters in the vegetative tissues of apple(Malus pumila mill.).Planta210:232-240.); A large number of heterologous proteins or metabolites accumulate in plants, breaking the metabolic balance of plants, not Conducive to plant growth (Robinson DJ (1996) Environmental risk assessment of releases of transgenic plants containing virus-derived inserts. Transgenic research5:359-362.); cause gene silencing or co-suppression (Kumpatla SP, Chandrasekharan MB, Iyer LM, Guofu L, Hall TC (1998) Genome intruder scanning and modulation systems and transgene silencing. Trends in Plant Science3:97-104; Mette MF, Aufsatz W, van der Winden J, Matzke MA, Matzke AJ (2000) Transcriptional silencing and promoter methylation triggered by double-stranded RNA.EMBO J19:5194-5201.); In addition, there are safety concerns of transgenic plants

Method used

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  • Floral organ specific expression promoter separated from Populus trichocarpa Torr.&Gray and application thereof
  • Floral organ specific expression promoter separated from Populus trichocarpa Torr.&Gray and application thereof
  • Floral organ specific expression promoter separated from Populus trichocarpa Torr.&Gray and application thereof

Examples

Experimental program
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Embodiment 1

[0050] Example 1 Cloning and sequence analysis of the PtAP1-2 gene promoter of Populus trichocarpa

[0051] According to the operating procedure of the Plant Genomic DNA Extraction Kit of Beijing Tiangen Company, the genomic DNA was extracted from the leaves of Populus trichocarpa clone and identified by 1.2% agarose gel electrophoresis. Primers were designed according to the 5' flanking sequence of Populus trichocarpa AP1-2 gene:

[0052] Upstream primer 5'-A GAGCTC TGCAACACTATTAATCAATTTATC-3',

[0053] Downstream primer 5'-A GGTACC CTCTTCTCTCTCTCTCTAAATG-3';

[0054] SacI and KpnI restriction sites were added to the 5' ends of the upstream and downstream primers, respectively. The 20 μL PCR reaction system includes: 2 μL (100 ng) Populus trichocarpa genomic DNA, 2 μL 10×LA PCR buffer, 1.6 μL (2.5 mmol / L) dNTP, 0.4 μL 10pm / μL upstream primer, 0.4 μL 10pm / μL downstream primer, 0.2 μL LA Taq DNA polymerase (5U / UL), 13.4μL ddH 2 O. The cycle conditions of the PCR reacti...

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Abstract

The invention discloses a floral organ specific expression promoter separated from Populus trichocarpa Torr.&Gray and application thereof, belonging to separation and application of a promoter. The nucleotide sequence of the promoter is shown as SEQ ID NO.1. The invention also discloses a recombinant plant expression vector and recombinant host cell containing the floral organ specific expression promoter. Functional verification tests discover that a GUS gene driven by the promoter only performs specific expression in sepals and petals of tobacco, and no GUS expression activity exists in roots, stems, leaves, stamens, carpels and other tissues of the tobacco, thus indicating that the promoter separated from Populus trichocarpa Torr.&Gray is the floral organ specific expression promoter. The invention further discloses application of the floral organ specific expression promoter in the aspects of improving plant traits, cultivating new plant species, studying floral organ development and regulation mechanisms of plants and the like.

Description

technical field [0001] The present invention relates to a promoter isolated from plants, in particular to a flower organ-specific expression promoter isolated from Populus trichocarpa Torr. & Gray, and the present invention also relates to a promoter containing the flower organ-specific expression promoter Recombinant plant expression vectors and host cells, the present invention further relates to their application in the study of flower organ gene expression patterns or transcriptional regulation mechanisms, improvement of plant traits, cultivation of new plant varieties, etc., belonging to the isolation of plant tissue or organ-specific expression promoters and its fields of application. Background technique [0002] The promoter is a DNA sequence that RNA polymerase specifically recognizes and binds to, and is an important cis-acting element, generally located in the upstream region of the 5' end of the structural gene. Gene regulation in higher plants is mainly carried...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82C12N15/84C12N1/21C12N5/10A01H5/00
Inventor 安新民陈仲王佳李英叶梅霞季乐翔
Owner BEIJING FORESTRY UNIVERSITY
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