Plant nutrition storage protein (GmVSPB) gene

A nutrient storage protein and gene technology, applied in the field of genetic engineering, can solve problems such as no significant increase in conversion rate and few breakthroughs

Inactive Publication Date: 2015-09-02
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
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  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported at home and abroad to use different soybean tissues as explants to successfully regenerate plants through organogenesis or embryogenesis and protoplast culture, but there are few breakthroughs, that is, the transformation rate has not been significantly improved.

Method used

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  • Plant nutrition storage protein (GmVSPB) gene
  • Plant nutrition storage protein (GmVSPB) gene
  • Plant nutrition storage protein (GmVSPB) gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] one. Experimental Materials and Reagents

[0027] 1. Plant material - soybean variety: Dongnong 50 with high regeneration rate, from Wu Xiaoxia's laboratory.

[0028] 2. Strains and vectors: Escherichia coli E.coli DH5α; Agrobacterium tumefacieus LBA4404; vectors pCAMBIA3300, pBI121; pMD18-T vectors were purchased from TaKaRa Company.

[0029] two. Main experimental reagents

[0030] CLONTECH SMARTer TM PCR cDNA Synthesis Kit and CLONTECH PCR-Select TM cDNA Subtraction Kit was purchased from CLONTECH; RNAiso Plus, pMD18-T, LA Taq DNA polymerase DL2000, DL8000, DL15000 were purchased from TaKaRa; restriction enzymes EcoRI, HindIII, SacI, SmaI were purchased from Fermantas; SYBY Green I Fluorescence quantitative kits were purchased from Dalian Bao Biological Company; primer synthesis and sequencing were completed by Beijing BGI.

[0031] three. experimental method

[0032] (1) Suppression subtractive hybridization

[0033] 1. Obtaining soybean cotyled...

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Abstract

A plant nutrition storage protein (GmVSPB) gene belongs to the technical field of genetic engineering and is characterized in that soybean Dongnong50 cotyledonary node is used as a material in suppression subtractive hybridization of differential gene when induced by cytokinin 6-BA so as to obtain a regeneration-associated gene and carry out gene cloning; a plant is transformed by agrobacterium-mediated transformation, changes of the transgenic plant are observed, gene functions are preliminarily analyzed by a quantitative RT-PCR method, and the influence of the cytokinin on regeneration is researched; starting from the influence of cotyledonary node organogenesis regeneration binding hormone on expression of soybean regeneration pathway associated transcription factor gene, effects of regenerating gene in a regeneration system are analyzed, and a soybean plant nutrition storage protein (GmVSPB) gene (Seq ID No:1) is found out.

Description

technical field [0001] The invention relates to a plant nutrient storage protein (GmVSPB) gene, which belongs to the technical field of genetic engineering. Background technique [0002] Soybean is an important grain and oil crop in my country. However, facing the shrinking arable land area and increasingly prominent food security problems, it is necessary to accelerate the pace of molecular breeding research in order to get rid of the shortage of soybean supply in my country and solve the severe situation of soybean dependence on imports. Strive to substantially increase the unit yield and total yield of soybeans in the short term. Biotechnology is the main means of molecular breeding, and a key guarantee for the application of biotechnology in soybean breeding is an efficient and stable regeneration system. Because soybean regeneration is affected by genotype, only a few soybean varieties have relatively stable regeneration systems, thus hindering the application of biotec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415
Inventor 苏安玉武小霞孙晶
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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