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A pumpkin mosaic virus recombinant vector carrying enhanced green fluorescent protein reporter gene and its construction method

A technology of green fluorescent protein and pumpkin mosaic virus, which is applied in the field of genetic engineering, can solve the problems of cucurbit crop planting and production threats, plant leaf shrinkage, serious diseases, etc.

Active Publication Date: 2020-10-23
ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Due to the high seed transmission rate of the virus, and the infection of the virus can cause mosaic and shrinkage of the leaves of the plant, and the disease is more serious during the flowering and fruiting stage, which poses a great threat to the planting and production of Cucurbitaceae crops.

Method used

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  • A pumpkin mosaic virus recombinant vector carrying enhanced green fluorescent protein reporter gene and its construction method
  • A pumpkin mosaic virus recombinant vector carrying enhanced green fluorescent protein reporter gene and its construction method
  • A pumpkin mosaic virus recombinant vector carrying enhanced green fluorescent protein reporter gene and its construction method

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Embodiment Construction

[0027] The technical solutions of the present invention will be clearly and completely described below in conjunction with the accompanying drawings and embodiments. Apparently, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0028] A pumpkin mosaic virus recombinant vector carrying an enhanced green fluorescent protein reporter gene. Its preservation number is CGMCC No. 13940, and its classification is named: Pumpkin mosaic virus. It was managed by China Microbial Culture Collection on November 22, 2018 The Committee's General Microbiology Center is responsible for preservation, and the detailed address is No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0029] A method for constructing a pumpkin mosaic ...

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Abstract

The invention relates to a SqMV (Squash Mosaic Virus) recombinant vector carrying an EGFP (Enhanced Green Fluorescent Protein) reporter gene and a construction method thereof. Infectious clone pSqMV-RNA2 corresponding to a SqMV genome RNA2 is used as a template, the EGFP reporter gene is inserted between MP and LCP to obtain the pSqMV-RNA2-EGFP recombinant vector; the vector can be inoculated to melons together with infectious clone pSqMV-RNA1 corresponding to a SqMV genome RNA1 by an agrobacterium co-infiltration method so as to obtain a SqMV capable of expressing the EGFP reporter gene. Theinvention provides a novel method and means for researching the infection process of the virus and disclosing a seed-borne mechanism of the virus and also specially provides a new tool for expressionof other exogenous genes or obtaining of a great quantity of application polypeptide.

Description

technical field [0001] The invention relates to a pumpkin mosaic virus recombinant vector carrying an enhanced green fluorescent protein reporter gene and a construction method thereof, belonging to the field of genetic engineering. Background technique [0002] An infectious clone of a virus refers to an infectious cDNA or an infectious RNA obtained by in vitro transcription. The acquisition of virus-infectious clones enables RNA viruses to apply genetic engineering methods at the DNA level to study the functions of viral genes and the pathogenic mechanism of viruses, and the insertion and co-expression of reporter genes in infectious clones can be more intuitively observed And record the infection process of the virus, and promote the research on virus pathogenicity. At present, green fluorescent protein (Greenfluorescent protein, GFP) is often used as a reporter gene. GFP is a light-emitting protein discovered in Aequorea victoria in 1962, consisting of 238 amino acids....

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/86C12N15/65C12N15/66
CPCC12N15/65C12N15/66C12N15/86C12N2770/00043
Inventor 刘莉铭古勤生
Owner ZHENGZHOU FRUIT RES INST CHINESE ACADEMY OF AGRI SCI