Vegetable lectin gene

A lectin and gene technology, applied in the fields of plant genetic improvement, plant peptide, genetic engineering, etc.

Inactive Publication Date: 2004-08-18
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although in the in vitro insect resistance test, ACA has shown a good anti-aphid effect, but whether it has the same function in transgenic plants remains to be confirmed experimentally

Method used

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  • Vegetable lectin gene
  • Vegetable lectin gene
  • Vegetable lectin gene

Examples

Experimental program
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Effect test

Embodiment

[0016] 1. Extraction of total DNA from amaranth Amaranthus caudatus

[0017] The plant material was amaranth (Amaranthus caudatus) provided by the Regional Plant Introduction Station of Iowa State University, USA. For the DNA extraction method, refer to literature 5. Take 200mg of amaranth leaves or mature seeds, grind with liquid nitrogen, add 1ml of ice-cold DNA extraction buffer [0.35M glucose, 0.1Mtris-Hcl (PH8.0), 0.005M Na2-EDTA (PH8.0), 2% (w / v) PVP, 0.1% (W / V) diethylthiocabarmic acid (DIECA), 0.2% (w / v) mercaptoethanol, 0.5% Triton-X100], after mixing thoroughly, centrifuge at 2700Xg for 10 minutes at 4°C , to the supernatant. Add 500ul nuclei lysis buffer [0.1M Tris-Hcl (PH8.0), 1.4M NaCl, 0.02M Na2-EDTA (PH8.0), 2% (W / V) CTAB, 2% (W / V) PVP, 0.1% (W / V) DIECA, 0.2% (W / V) mercaptoethanol, 0.5% Triton-X100], resuspend the precipitate, and place in a water bath at 65°C for 30 minutes. Add an equal volume of chloroform-isoamyl alcohol (24:1) for extraction, centrifug...

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Abstract

The clone of Amaranthus caudatus agglutinin (ACA) containing intron from Amaranthus caudatus and the gene sequence for coding ACA protein are disclosed. The said gene sequence is converted to efficient and stable microbe expression carrier for synthesizing a lot of ACA protein with microbes. The efficient expression of ACA gene in transgenic plant can make the said transgenic plant have high resistance to aphis and high nutritive quality.

Description

Technical field: [0001] The invention relates to the fields of plant insect-resistant genetic engineering and biochemistry. Specifically, the present invention relates to a newly cloned amaranth ACA gene and a recombinant plasmid expressed in plants, which can be transformed into plants to construct transgenic plants with aphid-resistant properties; meanwhile, by constructing a microbial expression vector for efficient and stable expression of the ACA gene , ACA protein can be obtained by using microorganisms. Background technique: [0002] ACA (Amaranthus caudatus agglutinin) is the seed of amaranth with high nutritional value in South America [1] A lectin present in , is a storage protein that provides nutrients during seed germination. This protein is only synthesized in large quantities when the endosperm is formed, and is not synthesized in the vegetative organs. The amino acid sequence of ACA protein has been reported in GenBank (Accession No.g2781234). The experim...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/415C12N15/29
CPCC07K14/415
Inventor 田颖川周永刚
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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