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Cloning and application of maize phosphoenolpyruvate carboxykinase gene promoter

A technology of pyruvate carboxykinase and phosphoenol, applied in the field of bioengineering breeding of crops

Inactive Publication Date: 2012-06-27
SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] According to the existing data, the cloning, analysis and utilization of the maize phosphoenolpyruvate carboxykinase gene promoter have not been reported.

Method used

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  • Cloning and application of maize phosphoenolpyruvate carboxykinase gene promoter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: Transformation of PZmPEPCK::Bt toxin gene to create corn insect-resistant inbred lines and its application

[0041] 1) The establishment of the receptor system is based on the backbone inbred lines used in agricultural production in my country, such as Zheng 58, Chang 7-2, DH4866 and other inbred seeds. Clustered buds were induced from the shoot tips in vitro, and the clustered buds were used as receptors for genetic transformation. The media used are:

[0042] Seed germination medium: KNO 3 1900mg / l, NH4NO 3 1650mg / l, CaCl 2 2H 2 O 440mg / l, MgSO 4 ·7H 2 O 370mg / l, KH 2 PO 4 ·H 2 O 170mg / l, FeSO 4 ·7H 2 O 27.8mg / l, ZnSO 4 ·7H 2 O 10mg / l, MnSO 4 4H 2 O 22.3 mg / l, H 3 BO 3 10mg / l, KI0.83mg / l, Na 2 MoO 4 2H 2 O 0.5mg / l, CuSO 4 ·5H 2 O0.025mg / l, CoCl 2 ·6H 2 O 0.025mg / l, Thiamine Hydrochloride 10.0mg / l, Pyridoxine Hydrochloride 1.0mg / l, Niacin 1.0mg / l, Glycine 2.0mg / l, Inositol 100.0mg / l, Biotin 0.05mg / l , casein hydrolyzate 500mg / l, sucros...

Embodiment 2

[0056] Example 2: Transformation of PZmPEPCK::TPT1 gene to create corn breeding materials with high light efficiency

[0057] The sugar / phosphorus transport between the plant cytoplasm and the chloroplast stroma is mainly responsible for the triose-phosphate / phosphate translocator (TPT), which can transport phosphorus from the cytoplasm to the chloroplast. At the same time, etc. Moles of triosephosphate 3-PGA are transported from the chloroplast stroma into the cytoplasm. When the cytoplasmic phosphorus concentration decreased, the activity of TPT transporter was inhibited, which led to the accumulation of metabolites in chloroplasts and affected the photosynthetic rate of leaves. ZmTPT1 is a triose phosphate / phosphate transporter gene cloned from maize. It has high affinity for substrates. By constructing a fusion gene, the gene can be highly expressed in maize vascular bundle sheath cells, which is conducive to maintaining a high photosynthetic rate in maize leaves. This ex...

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Abstract

The invention discloses cloning and application of a maize phosphoenolpyruvate carboxykinase gene promoter. A method comprises the following steps of: cloning a phosphoenolpyruvate carboxykinase gene promoter from maize, fusing the cloned phosphoenolpyruvate carboxykinase gene promoter with a target gene coding area or a ribonucleic acid interference (RNAi) structure thereof to construct a plant expression structure, and introducing a recombinant gene into a plant cell by transgenic technology to generate a transgenic plant; and selecting the target character of the transgenic plant to obtainthe transgenic plant of which the stress resistance or economic character is improved obviously or descendants thereof, so that new germplasm and species which have application prospects in plant breeding are created.

Description

technical field [0001] The invention belongs to the field of bioengineering breeding of crops, and specifically relates to a scheme and application for changing plant traits by constructing fusion genes and transgenes. Background technique [0002] The development of plant genetic engineering relies on the application of promoters with different properties. At present, the commonly used promoters in plant genetic engineering mainly include constitutive promoters and inducible promoters, such as cauliflower mosaic virus (CaMV) 35S promoter, nopaline synthase gene Nos promoter, quasi The A. thaliana Rd29A promoter, the maize Ubi1 promoter and the rice actin gene Act1 promoter commonly used in monocotyledonous transgenic plants. Constitutive promoters have the characteristics of continuous and constant expression of driving genes in various tissues, and can produce a large number of heterologous proteins, which is of great significance in the cultivation of insect-resistant an...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/00
Inventor 张举仁李朝霞李坤朋张广峰
Owner SHANDONG UNIV
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