Method for culturing transgenic alfalfa with high sulfur-containing amino acid content and special materials thereof

A technology of transgenic alfalfa and amino acids, applied in the direction of botany equipment and methods, applications, transferases, etc., can solve the problem of low content of sulfur-containing amino acids in proteins

Active Publication Date: 2013-07-24
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although alfalfa is high in protein, its protein is low in sulfur-containing amino acids

Method used

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  • Method for culturing transgenic alfalfa with high sulfur-containing amino acid content and special materials thereof
  • Method for culturing transgenic alfalfa with high sulfur-containing amino acid content and special materials thereof
  • Method for culturing transgenic alfalfa with high sulfur-containing amino acid content and special materials thereof

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Experimental program
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Effect test

Embodiment 1

[0060] Example 1. Utilizing the gene encoding aspartokinase and the gene encoding adenosine sulfate reductase to cultivate transgenic alfalfa with altered amino acid content

[0061] 1. Construction of the expression vector pCAMBIAK-APR for the gene encoding aspartate kinase and the gene encoding adenosine sulfate reductase

[0062] 1. Construction of the intermediate vector pOSB108-AK containing the expression cassette of the gene encoding aspartate kinase

[0063] The forward primer is 5'-CTCGAGTCTGAAATTGTTGTCTCCAA-3' and the reverse primer is 5'-TCTAGATTACTCAAAACAAATTACTAT-3'. The first to sixth positions of the forward primer from the 5' end are XhoI restriction sites, and the reverse primer is from the 5' end The 1st to 6th positions are the XbaI restriction sites, the aspartokinase coding gene (AK gene) is amplified from Escherichia coli K12, and the PCR amplification product is recovered. The PCR amplification products were sequenced. The sequencing result shows that ...

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Abstract

The invention discloses a method for culturing a transgenic alfalfa with high sulfur-containing amino acid content and special materials thereof. The method for culturing transgenic alfalfa with changed amino acid content provided by the invention introduces aspartokinase encoding gene and adenylylsulfate reductase encoding gene into receptor alfalfa to obtain transgenic alfalfa with amino acid content changed. The aspartokinase is a protein shown as 65-512th site in an amino acid sequence SEQ ID No.2; and the adenylylsulfate reductase is a protein shown as 65-330th in an amino acid sequence SEQ ID No.4. The methionine content of the transgenic alfalfa with changed amino acid content cultivated by the invention is significantly higher than namely 1.5 times of that of the receptor alfalfa of an untransformed Baoding alfalfa (wild type); cysteine content of a transgenic pCAMBIAK-APR line is significantly higher than namely 1.2 times that of the wild type; aspartic acid content of a transgenic pCAMBIAK-APR line is significantly higher than namely 1.4 times that of the wild type; and lysine content of a transgenic pCAMBIAK-APR line is significantly higher than namely 1.5 times that of the wild type.

Description

technical field [0001] The invention relates to a method for cultivating transgenic alfalfa high in sulfur-containing amino acids and special materials thereof. Background technique [0002] Alfalfa is a general term for plants of the genus Medicago. It is a perennial flowering plant, the most famous of which is Medicago sativa L. as pasture. Alfalfa is a very important perennial wild leguminous forage in the genus Medicago, mainly distributed in China, Mongolia, Russia, Pakistan, India, Turkey and other countries. my country's alfalfa species are rich in resources, mainly distributed in the northeast, north and northwest regions of China. Compared with other species, alfalfa has outstanding characteristics such as strong stress resistance, cold resistance, drought resistance, barren resistance and long life span. It is suitable for planting in arid and cold areas and is the first choice for building high-yield and high-quality artificial grassland in northern my country. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00C12N9/12C12N9/02C12N15/62
Inventor 王涛仝宗永董江丽
Owner CHINA AGRI UNIV
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