Ammopiptanthus mongolicus cold resistant gene AmGS

A kind of technology of sand holly and gene, applied in the field of cold-resistant gene AmGS of sand holly, can solve the problems of cold-resistant gene cloning and transformation that have not been successfully reported.

Inactive Publication Date: 2012-07-18
TAISHAN RES INST OF FORESTRY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there have been no successful reports on the cloning and transformation of cold resistance genes in woody plants

Method used

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  • Ammopiptanthus mongolicus cold resistant gene AmGS
  • Ammopiptanthus mongolicus cold resistant gene AmGS
  • Ammopiptanthus mongolicus cold resistant gene AmGS

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0007] (1) The in vitro regeneration system of woody plants is used as the transformation material, and genetic transformation is carried out by means of Agrobacterium dipping, pollen tube passage, gene gun and other methods.

[0008] (2) Conversion operation steps

[0009] 1. Agrobacterium transformation method

[0010] (1) The pCAMBIA2300-AmGS expression vector was transformed into Agrobacterium tumefaciens LBA4404 by triparental hybridization method for the Agrobacterium dipping transformation experiment.

[0011] (2) Bacterial solution preparation

[0012] 1) Use an inoculation loop to scrape the surface of the frozen culture of Agrobacterium strains containing recombinant plasmids, streak on the above-mentioned YEP plate containing the corresponding antibiotics, and place it in a constant temperature incubator at 28°C for 2 days, until colonies grow out on the plate It can be used for liquid culture. Plates with colonies were stored in a 4°C refrigerator. After one mo...

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Abstract

The invention provides an ammopiptanthus mongolicus cold resistant gene AmGS and relates to an endangered evergreen broad leaf woody plant considered as the second grade protection in the country and derived from the ancient tertiary period in the northwest desert region in China. According to molecular cloning the gene, the EST segment of the ammopiptanthus mongolicus cold resistant gene with low temperature induced up regulated expression is obtained through low temperature induction and differential expression analysis, and the full length cDNA sequence of the gene is obtained through RACE amplification. The AmGS comprises a complete ORF region; the ORF full length comprises 987 nucleotides; 328 amino acids and a termination codon (TAA) are coded; a plant expression vector pCAMBIA2300-AmGS of the AmGS gene is established; and the pCAMBIA2300-AmGS is used for transforming woody plants such as photinia fraseri and apricot trees, so that transgenic plants with improved cold resistance are obtained.

Description

1. Technical field [0001] The present invention relates to a national second-class endangered protected plant originating from the ancient Tertiary in the desert area of ​​northwest my country, which is also the molecule of the cold resistance gene AmGS of the evergreen broad-leaved woody plant Ilex chinensis preserved in the desert area of ​​central Asia so far Cloning and genetic transformation. Through low-temperature induction and differential expression analysis, the EST fragment of the antifreeze gene of A. chinensis was obtained, and then the full-length cDNA sequence of the antifreeze gene AmGS of A. chinensis was obtained by RACE amplification, which was registered in the gene bank. , registered as DQ519359. AmGS contains a complete ORF region, the full length of ORF is 987 nucleotides, encoding 328 amino acids and a stop codon (TAA). Subsequently, the eukaryotic expression vector pCAMBIA2300-AmGS of the cold resistance gene AmGS of Ilex chinensis was constructed and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C12N15/82C12N15/11A01H5/00
Inventor 冯殿齐王斌刘静宋键曹鹏秀翁曼丽罗磊
Owner TAISHAN RES INST OF FORESTRY
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