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Plants lamina vascular bundle specificity expressive promotor and application thereof

A technology for plant leaves and plant expression vectors, applied to plant expression vectors, the application of the above promoter in plant transgenic engineering, and the field of plant vascular bundle-specific expression promoters can solve the problem of difficult to achieve directional transformation of plant traits and plant growth. Waste of resources, not showing the specificity of time and space, etc., to achieve the effect of improving resistance, reducing impact, and increasing effect

Inactive Publication Date: 2008-09-17
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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AI Technical Summary

Problems solved by technology

The promoter is necessary for gene expression and determines the space, time, and expression level of exogenous gene expression. It is a crucial factor in biotransgenic engineering and an important factor related to the safety of biotransgenic. It was widely used at the earliest The biological transgene promoters are some constitutive promoters, such as the 35S promoter derived from CaMV, the nopaline synthase gene promoter Nos in the T-DNA region of the Ti plasmid of Agrobacterium tumefaciens, the octopine synthase gene promoter Ocs, Constitutive promoters are characterized by starting gene expression in all tissues, which are persistent, do not show spatiotemporal specificity, are difficult to achieve the purpose of directional transformation of plant traits, and often produce some unwanted results or even negative effects. In addition The continuous expression of constitutive promoters in time and space also causes a waste of plant growth resources

Method used

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  • Plants lamina vascular bundle specificity expressive promotor and application thereof
  • Plants lamina vascular bundle specificity expressive promotor and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Example 1: Obtaining of the promoter Op containing restriction sites

[0029] Step 1. Design of primers

[0030] According to the whole genome sequence of rice variety Nipponbare (Oryza sativa L cv. Nipponbare) provided by NCBI, the amplification primers were designed according to the 1380 bp sequence upstream of the start codon of rice OsBTF3 gene, and the primers were designed according to the characteristics of the selected vector and target gene In this embodiment, the rice binary expression vector pCAMBIA1301 (from CAMBIA, openly used carrier, Molecular Plant Disease Group, State Key Laboratory of Plant Disease and Pest Biology, Institute of Plant Protection, Chinese Academy of Agricultural Sciences has been preserved, public can be obtained from this laboratory) as an example, the target gene is the GUS gene, and the specifically designed primers are: the 5' end of the forward primer Op1 has a BglII restriction site, and the 5' end of the reverse primer Op2 has an...

experiment example 2

[0039] Experimental example 2: Using the Op promoter to drive the expression of GUS reporter gene in rice

[0040] Step 1 Construction of plant expression vector and transformation of Agrobacterium

[0041] The pMD18-T-Op obtained in Example 1 was digested with Bgl II and EcoR I, and after recovery, it was digested with the Bgl II and EcoR I of pCAMBIA1301 to digest the vector part, and connected with T4 ligase (purchased from Takara Biological Company) to obtain the promoter The plant expression vector pCAM-Op ( figure 2 ), using the freeze-thaw method to transfer the expression vector into Agrobacterium tumefaciens (Agrobacterium tumefaciens) EHA105 (the Molecular Plant Disease Group of the State Key Laboratory of Plant Disease and Pest Biology, Institute of Plant Protection, Chinese Academy of Agricultural Sciences has preserved it, and the public can get it from this laboratory. ), extract the positive plasmid, carry out enzyme digestion verification with BglII and EcoR ...

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Abstract

The invention relates to the plant gene engineering field, particularly a plant leaf promoter conferring vascular-specific expression, a plant expression vector comprising the promoter and a transformant. The invention also relates to the application of the promoter in the plant gene engineering. The nucleotide sequence of the plant leaf promoter conferring vascular-specific expression is shown as SEQ ID No:1. The inventor subjects the plant expression vector with the promoter DNA sequence and a GUS gene to genetic transformation mediated through agrobacterium tumfaciens to obtain transgenic plants. According to identification by immunocytochemistry, the GUS gene only confers vascular -specific expression on rice leaves, therefore, it is supposed that the promoter provided in the invention can drive extrinsic stress-resistant genes to confer vascular -specific expression on plant leaves, so that the promoter can be directionally used for cultivate new transgenic plants with high safety.

Description

technical field [0001] The present invention relates to the field of plant genetic engineering, in particular to a promoter specifically expressed in plant vascular bundles, a plant expression vector containing the promoter, and a transformant of the promoter. application in engineering. Background technique [0002] The promoter is located in the upstream region of the 5' end of the structural gene, which can be recognized by RNA polymerase and bind to it, thereby initiating the transcription of a DNA sequence of the gene. The promoter is necessary for gene expression and determines the space, time, and expression level of exogenous gene expression. It is a crucial factor in biotransgenic engineering and an important factor related to the safety of biotransgenic. It was widely used at the earliest The biological transgene promoters are some constitutive promoters, such as the 35S promoter derived from CaMV, the nopaline synthase gene promoter Nos in the T-DNA region of the...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82C12N5/10A01H1/00A01H5/00
Inventor 何晨阳吴茂森柏亚男
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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