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Plant low-phosphorus response regulation and control unit and expression vector construction technology

A plant expression vector, plant technology, applied in plant products, plant genetic improvement, recombinant DNA technology, etc., to improve absorption and utilization efficiency, reduce resource waste and environmental pollution, and reduce the use of phosphate fertilizers.

Inactive Publication Date: 2014-12-03
NANKAI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After searching the literature, no literature or patent reports have been found that PHR1 gene and SIZ1 gene are constructed as independent regulatory units in the same vector to transform plants to improve plant phosphorus uptake and utilization efficiency

Method used

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  • Plant low-phosphorus response regulation and control unit and expression vector construction technology
  • Plant low-phosphorus response regulation and control unit and expression vector construction technology
  • Plant low-phosphorus response regulation and control unit and expression vector construction technology

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Cloning of the Arabidopsis SIZ1 gene. Including the following steps:

[0035] (1) Arabidopsis total RNA was extracted by TRIZOL reagent method, and its cDNA was obtained by reverse transcription.

[0036] (2) Using cDNA as a template PCR clone to obtain the SIZ1 gene ( figure 2 ), clone it into pMD19-T simple vector, construct the vector pMD19-T-SIZ1 for sequencing, and the sequence is correct. The primers used in PCR were designed according to the Arabidopsis AtSIZ1 gene sequence:

[0037] P1: 5′- GAATTC ATGGATTTGGAAGCTAATTGTAAGGAAAAAC-3′

[0038] P2: 5′- ACTAGT CAAATCTTGTTTAAACTCCGGTGTCT-3′

[0039] An EcoR I restriction site is added to the upstream primer P1, and a Spe I restriction site is added to the downstream primer P2, both of which are underlined.

Embodiment 2

[0041] Construction of a low phosphorus response regulatory unit expression plant vector. Including the following steps:

[0042] (1) Construction of pMD19-T-35S vector.

[0043] The 35S promoter ( image 3 ) into pMD19-T simple vector to construct vector pMD19-T-35S. The PCR primers used were designed according to the DNA sequence of the 35S promoter:

[0044] P3: 5′- GAATTCACTAGT CCCACAGATGGTTAG-3′

[0045] P4: 5′- GAATTC CGTGTTTCTCCAAATGAAAT-3′

[0046] The upstream primer P3 adds EcoR I and Spe I restriction sites, and the downstream primer P4 adds an EcoR I restriction site, both of which are underlined.

[0047] (2) Construction of pMD19-T-35S-SIZ1 vector.

[0048] Using pMD19-T-SIZ1 as the basic vector, the 35S promoter was recovered from the vector pMD19-T-35S after being digested with EcoR I, inserted into the vector pMD19-T-SIZ1, and identified by PCR and enzyme digestion ( Figure 4 ) and identification of its insertion forward and reverse ( Figure 5 ),...

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Abstract

The invention describes a construction technology of an expression vector pX6-PHR1-35S-SIZ1 of a plant low-phosphorus response regulation and control unit PSS (PHR1-35S-SIZ1). The construction method of the expression carrier comprises: cloning and obtain SIZ1 gene from arabidopis thaliana, inserting 35S promoter into the upstream of SIZ1 gene, so as to construct a 35S-SIZ1 expression unit, inserting the expression unit into a plant expression vector pX6 to construct a vector pX6-35S-SIZ1, then inserting PHR1 gene into the vector, so as to obtain pX6-PHR1-35S-SIZ1. The expression vector pX6-PHR1-35S-SIZ1 is used to transform agrobacterium GV3101 so as to obtain an engineering agrobacterium strain NK-314 containing both PHR1 gene and SIZ1 gene.

Description

technical field [0001] The invention relates to a plant low phosphorus response control unit and expression carrier construction technology, which belongs to the field of biological technology. Background technique [0002] Phosphorus is one of the indispensable nutrient elements for plant growth and development. It is not only a component of ATP, nucleic acid, and phospholipid molecules in organisms, but also plays a key role in energy conversion and metabolism regulation. However, phosphorus is mainly in the form of H 2 PO 4 - and HPO 4 2- The soluble form is absorbed by plants, and this part of phosphorus is called "available phosphorus". In soil, although the phosphorus content is high, most of the phosphorus is absorbed by Ca 2+ , Fe 2+ 、Al 3+ Phosphate transport in plants. Phosphate transport in plants. Plant and Soil, 2003, 248(1- 2): 71-83). [0003] PHR1 is a kind of conserved MYB transcription factor, which can bind to the promoters of some phosphorus sta...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/52C12N15/63C12N15/74C12N15/82A01H5/00C12R1/01
Inventor 李明刚郝军凤乔坤艳马超马百成王宇李宠张耀方马志华姬艳丽董震屠培培李晓丹
Owner NANKAI UNIV
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