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Low-virulent vaccine K genom sequence of tomato mosaic

A technology of tomato mosaic virus and attenuated vaccine, applied in the direction of genetic engineering, plant genetic improvement, application, etc.

Inactive Publication Date: 2004-05-19
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

Its physical and chemical properties, changes in double-stranded RNA in susceptible tissues, immune proteins, and RNA in vitro translation products have been studied, but there is no clear report on its weakening mechanism

Method used

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  • Low-virulent vaccine K genom sequence of tomato mosaic
  • Low-virulent vaccine K genom sequence of tomato mosaic
  • Low-virulent vaccine K genom sequence of tomato mosaic

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Experimental program
Comparison scheme
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Embodiment

[0025] 1. The point mutation makes the genome cDNA replica enzyme coding gene of tomato mosaic virus form a milkstone mutation (TGA) at 2670-2672 nucleotides, and the motor protein coding gene forms an ocher mutation (TAA) at 5632-5634 nucleotides. Point mutation can use PCR (polymerase chain reaction) to introduce the mutation site into the primer. The method is as follows:

[0026] (1) Mutation primer design:

[0027] The upstream and downstream (5' and 3') primers must include the mutation site, and after annealing, the two are completely paired to form a double strand; the replicase 5' mutation primer sequence is: 5-GATGATCAGA TGA AGAGCTAATG-3', 3' Mutation Primer: 5'-CATTAGCTCT TCA TCTGATCATC-3'. Motor protein 5' mutation primer: 5'-AGTTTAAAAAGA GTTTAATAATTTGATTAAAGATTAAGC-3', 3'

[0028] Primer: 5'-GCTTAATCTTTAATCAAATTATTAAACTCTTTTTAAACT-3';

[0029] The 5' end of the primer cannot be phosphorylated, but the primer must be purified by polynucleotide liquid chromato...

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Abstract

The strong genom separated and cloned from tomato mosaic is modified into low-virulent vaccine K genom by point mutation. The nucleotide sequence of its coded replicase and motor protein has the mutation, so it can be used to modify the plant virus genom and prepare genetically engineered vaccine for effectively preventing and treating the viral diseases of plant.

Description

technical field [0001] The invention relates to the field of plant virus genetic engineering, in particular to the full sequence of a tomato mosaic virus attenuated vaccine genome of the tobacco mosaic virus genus and the weakened mutation mode. Background technique [0002] Plant virus attenuated vaccines play an important role in the prevention and treatment of plant virus diseases. They can survive and reproduce in plants without harming the normal growth, flowering and seed production of plants, and have a defensive effect on plants infected by attacking viruses. The phenomenon of symbiosis and host protection is the common feature of most attenuated plant viruses and is universal. Kunkel, Grant, Posnett and Todd have successively proposed the idea of ​​using plant natural attenuated virus isolates to prevent strong virus infection, and have done protection experiments with naturally isolated attenuated viruses in the study of fruit tree viruses. However, because these ...

Claims

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Application Information

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IPC IPC(8): C12N7/04C12N15/33
Inventor 邱并生杨恭田波
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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