Yeast genetically engineered strain expressing hrp gene and constructing method thereof

A technology of genetically engineered strains and construction methods, applied in the field of yeast genetically engineered strains and their construction, can solve the problems of small harvest, complicated process, and inability to meet market needs, etc.

A technology of genetically engineered strains and construction methods, applied in the field of yeast genetically engineered strains and their construction, can solve the problems of small harvest, complicated process, and inability to meet market needs, etc.

CN1454989AInactive Publication Date: 2003-11-12WUCAI FINE CHEM JIANGSU
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  • Yeast genetically engineered strain expressing hrp gene and constructing method thereof
  • Yeast genetically engineered strain expressing hrp gene and constructing method thereof
  • Yeast genetically engineered strain expressing hrp gene and constructing method thereof

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Experimental program
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Embodiment 1

[0108] Construction method of genetic engineering strain HrpZ

[0109] 1. Genomic DNA extraction and hrpZ gene isolation and cloning

[0110] 1. Genomic DNA Extraction

[0111] A strain of Pseudomonas syringae was isolated and identified from the soil, and the genomic DNA was extracted according to the instructions of the Genomic DNA Extraction Kit from QIAGEN Company.

[0112] 2. Isolation of hrpZ gene

[0113] The hrpZ gene was amplified and isolated by PCR using the genomic DNA of Pseudomonas syringae as a template, and the PCR primer sequences were as follows:

[0114] Forward: 5'-ATGCAGAGTCTCAGTCTTAACAGCA-3'

[0115] Reverse: 5'-AGTTCTCCGTCAGGCGGTTGTC-3'

[0116] PCR reaction conditions:

[0117] Pre-denaturation: 94°C for 1 minute

[0118] Denaturation: 92°C for 45 seconds

[0119] Refolding: 66°C for 45 seconds

[0120] Extension: 68°C for 2 minutes

[0121] Cycles: 30

[0122] Termination of extension: 68°C for 20 minutes

[0123] The PCR product was detecte...

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Abstract

The present invention relates to a yeast gene engineering strain capable of expressing hrp gene and its construction method. The name of said strain is Saccharomyces cerevisiae hrp Z, its number is CCTCC No:M203004. Its construction method includes: extraction of pseudomonas syringae genome DNA; using genome DNA as template PCR primer to make construction of hrp Z recombinant yeast gene engineering strain; hrpZ gene contained yeast conversion and induction. The gene engineering harpin protein produced by said strain can induce plant hypersensitive reaction, activate internal self-defense function of plant and promote plant growth. so that it can raise yield and quality of crops.

Description

technical field [0001] The invention relates to bioengineering, in particular to a yeast gene engineering strain expressing hrp gene and its construction method. Background technique [0002] The hrp gene cluster exists in a variety of plant pathogenic bacteria, and the Harpin protein encoded by it is a protein necessary for bacterial pathogenicity, but for non-host plants, it can cause hypersensitive responses of plants (Hypersensitive response), its Expressed by rapid, localized atrophy and death of the infected tissue, this limits the possibility of further spread of the pathogen. Hypersensitivity response is a kind of protective response that plants have, which is similar to the broad-spectrum immune response against pathogens and pests in higher animals. The Harpin protein itself has no killing or inhibiting effect on bacteria, viruses, fungi, nematodes, and insects. Instead, it binds to plant receptors and generates a certain signal to stimulate or induce the plant's ...

Claims

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Application Information

Patent Timeline
12 Nov 2003
Publication
CN1454989A
IPC
C12N1/19; C12N1/20; C12N5/10; C12N15/31; C12N15/70; C12N15/81; C12P21/00
Inventors
魏晨飞