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The Preliminary Research onβ-Conglycinin α Subunit Gene into Soybean

Author: LiMeng
Tutor: YuDeYue
School: Nanjing Agricultural College
Course: Crop Genetics and Breeding
Keywords: Soybean(Glyccine max (L.)Merr.) Sulfur amino acid α-subunit ofβ-conglycinin Agrobacterium-mediated Glufosinate
CLC: S565.1
Type: Master's thesis
Year: 2013
Downloads: 3
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Abstract

RNA interference(RNAi) is the post-transcriptional gene-silencing of the specfic gene induced by double-strand RNA.Soybean storage protein gene expression is regulated by both transcriptional and post-transcriptional processes.lt has been suggested that the health management and food processing of soybean protein could be improved by increasing glycinin content or decreasing P-conglycinin content.The total protein and fat content of soybean will not be changed.Thus it has become possible to get more information about the contribution to improving the sulfur amino acids score and the11S and7S regulatory mechanisms by RNAi.To identify the characteristic about the a subunit gene of β-conglycinin,the research took some studys on several respects:Firstly,After blast the sequence (GenBank No.AB051865) in Soybean Genome Database,two a-subunit copies were annotated as Glyma20g28650and Glyma20g28660in the Williams82genome sequence.They are all located in chromosome of NO.20with6exons and5itrons.The ORF section is no diversity which is composed of605-amino acid.Gm7Sa protein sequences showed striking similarity to homologous genes from Mt and relatively distant homology to Pt.There are a transmembrane region and protein signal peptide at the N-terminus of the protein in7Sa gene expression.The research analyzed the feature of Gm7Sa,and constructed the RNA interference vector by Gateway technology.Then the vector was get into the cell of the soybean by Agrobacterium-mediated genetic transformation.By the way,the sterilization time and sensitivity on the glufosinate of different genotypes of soybean was explored in order to improve the conversion rate.After repeatedly experiments,71soybean transgenic plants has got,with161seeds.Due to the impact of the bird damage,we totaly got65soybean transgenic plants after sowing in the field.We observed the change of the phenotype of the soybean transgenic plants.Through the smearing experimental results of bar test,identification of PCR,determination of copies of RT-PCR and so on,there are13nannong88-1,7Jack. The advanced generation strains with RNAi-Gm7Sa was selected by pedigree method.The seeds of the fine individual plant made the trip after harvest.Molecular detection and amino acid content assay of advanced generation soybean strains showed that the plant with low Gm7Sa gene expression had a high sulfur amino acids score.

The Preliminary Research onβ-Conglycinin α Subunit Gene into Soybean

Abstract

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