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Phosphorus transporter gene and preparation method thereof

A technology for transporting proteins and genes, applied in the fields of botanical equipment and methods, biochemical equipment and methods, genetic engineering, etc.

Inactive Publication Date: 2011-12-14
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, through literature search of the prior art, it is found that there is no description and report of the phosphorus transporter gene of Rosaceae plants so far

Method used

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  • Phosphorus transporter gene and preparation method thereof
  • Phosphorus transporter gene and preparation method thereof
  • Phosphorus transporter gene and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Cloning of Phosphorus Transporter Gene cDNA in Soybean Pear

[0030] Design primer pairs:

[0031] Upstream primer: SEQ ID NO:3

[0032] 5'-GTGCTNAATGCACTNGATGT-3'

[0033] Downstream primer: SEQ ID NO:4

[0034] 5'-CCTDGCHGGGAAAATCTCNGC-3'

[0035] In 0.230 g?L -1 NH 4 h 2 PO 4 The pear root treated for 24 h was used as plant material, and total RNA was extracted with RNAplant Plus Kit (purchased from TIANGEN Company), and the operation process was strictly in accordance with the extraction procedure in the manual. After being digested with DNase I (RNase Free, TIANGEN Company), 2 μg of total RNA was used for the synthesis of the first strand of cDNA for amplification of conserved region fragments (TIANScript cDNA First Strand Synthesis Kit, TIANGEN Company), and the reverse transcription primer was oligo(dT) 15, RNase A (DNAase and Protease-free, GenStar Company) was used to digest the first strand of cDNA used for amplification of the conserved ...

Embodiment 2

[0102] Example 2 Sequence Information and Homology Analysis of Soybean Phosphorus Transporter Gene

[0103] PcPht1 Gene nucleotide translation uses BioXM software, PcPht1 The transmembrane region analysis of gene-encoded proteins was performed using the TMHMM Server v. 2.0 program (http: / / www.cbs.dtu.dk / services / TMHMM). Secondary structure prediction and domain analysis were performed by PBIL LYON-GERLAND (http: / / npsa-pbil.ibcp.fr / cgi-bin / npsa_automat.pl / page= / NPSA / npsa_hnn.html) and InterPro (http: http: / / www.ebi.ac.uk / InterProScan) done. Subcellular localization was performed with the online software PSORT (http: / / psort.ims.utokyo.ac.jp / form.html). The amino acid sequence alignment was completed using DNAMAN software, and the Pht1 protein sequences of other plants used were from http: / / www.ncbi.nlm.nih.gov.

[0104] PcPht1 The gene-encoded protein is a protein with 538 amino acid residues:

[0105] MAREQLQVLN ALDVAKTQWY HFTAIIIAGM GFFTDAYDLF CISLVTKLLG RIYYHVEGAA ...

Embodiment 3

[0117] Example 3 Study on the Expression of Soybean Phosphorus Transporter Gene

[0118] When the pear seedlings grow to the size of 4 true leaves, select the plants with the same growth and starve for 24 h in deionized water, then place them in NH 4 h 2 PO 4 0.023, 0.230 or 2.30 g?L -1 The Hoagland nutrient solution was treated for 24 h, and the leaves and roots of different treatments were harvested and stored in a -70°C refrigerator for later use. Hoagland nutrient solution formula: 1.02 g?L -1 KNO 3 , 0.492 g?L -1 Ca(NO 3 ) 2 , 0.023, 0.230 or 2.30 g?L -1 NH 4 h 2 PO 4 , 0.49 g?L -1 MgSO 4 ?7H 2 O, 24.5 mg?L -1 Ferric citrate, 2.230 mg?L -1 MnSO 4 , 0.240 mg?L -1 CuSO 4 ?5H 2 O, 0.290 mg?L -1 ZnSO 4 ?7H 2 O, 1.860 mg?L -1 h 3 BO 3 , 0.035 mg?L -1 (NH 4 ) 6 Mo 7 o 24 ?4H 2 O, 0.028 mg?L -1 CoSO 4 ?H 2 O.

[0119] Total RNA was extracted with RNAplant plus Total Plant RNA Extraction Reagent (TIANGEN Company), and the operation...

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Abstract

The present invention relates to a phosphorus transporter gene, its encoded protein and a preparation method of the gene. The phosphorus transporter gene is derived from a Rosaceae plant, the phosphorus transporter gene is SEQ ID NO: 1, and its encoded protein is SEQ ID NO: 2; The phosphorus transporter gene can be cloned by the designed primers; the phosphorus transporter gene can be used to study the expression of the phosphorus transporter gene in Rosaceae plants, and under different phosphorus supply conditions, the gene can regulate its own expression level , to meet the needs of plants for phosphorus transport.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a phosphorus transporter gene expressed in Rosaceae plants and its clone. Background technique [0002] Phosphorus is an important nutrient element in the life process of plants, constituting 5% of the dry weight of plant cells. As a substance for energy transfer, phosphorus plays an important role in activating proteins in the body, regulating the entire metabolic process of plants, coordinating growth and stress resistance, etc. (Bucher M. Functional biology of plant phosphate uptake at root and mycorrhiza interfaces [J]. New Phytologist, 2007, 173(1): 11-26). Plants mainly absorb phosphorus in the soil in the form of inorganic phosphorus (Pi), and the strong adsorption of phosphorus by the soil makes the content of soluble inorganic phosphorus in the soil solution that can be absorbed by plants very low, usually less than 10 μmol L -1 (Raghothama K G. and Karthikeyan ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/10
Inventor 李慧丛郁常有宏蔺经
Owner JIANGSU ACAD OF AGRI SCI
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