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A cp gene of a latent virus of Vetch and its application

A latent virus and vetch technology, which is applied in the fields of application, genetic engineering, plant genetic improvement, etc., can solve the problems of undetected vetch latent virus, and achieve the effect of easy identification, simple operation and high sensitivity

Active Publication Date: 2022-05-06
JIANGSU ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, LAMP technology has been used to detect soybean mosaic virus (SMV) and bean pod mottle virus (BPMV), but there is no report on using LAMP technology to detect vetch latent virus.

Method used

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  • A cp gene of a latent virus of Vetch and its application
  • A cp gene of a latent virus of Vetch and its application
  • A cp gene of a latent virus of Vetch and its application

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Experimental program
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Effect test

Embodiment 1

[0028] 1. Test materials

[0029] The broad bean samples to be tested in the field were collected in Lishui District, Nanjing City, Jiangsu Province in September 2019. The samples were placed in a portable freezer with ice packs and brought back to the laboratory for RNA extraction. The extracted RNA was stored in a -80°C refrigerator for later use . The sample was confirmed to contain Vicia cryptic virus (VCV) by ordinary RT-PCR detection, and the CP gene sequence of VCV was connected into the pClone007 vector (purchased from Beijing Qingke Biotechnology Co., Ltd.) to construct pClone007-VCV -CP carrier was used as the experimental material.

[0030] 2. Design of main reagents and primer sequences

[0031] RNAprep pure Plant Total RNA Extraction Kit, FastKing one-step removal of genomic cDNA first-strand synthesis premix reagent were purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd., pClone007 Blunt Simple Vector Kit, DNA Gel Recovery Kit, PCR Master Mix Bo...

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Abstract

The invention discloses a CP gene of a vetch latent virus. The nucleotide sequence of the CP gene is shown in SEQ ID NO:1. The invention also discloses a recombinant vector comprising the CP gene, a pair of specific primers for amplifying the CP gene, a primer set for detecting the CP gene of the vetch latent virus, a detection kit for the vetto latent virus and A method for the detection of latent viruses in vetch for non-diagnostic purposes. The detection kit and its detection method of the present invention do not require a PCR instrument, are low in cost and simple in operation, and can amplify a limited amount of DNA to 1 million copies with high specificity and high efficiency within 1 hour, and its sensitivity is higher than that of PCR technology, and the response results are easily recognizable.

Description

technical field [0001] The invention belongs to the biological field, and in particular relates to a CP gene of a vetch latent virus and an application thereof. Background technique [0002] Vicia cryptic virus (VCV) is a rare double-stranded RNA (dsRNA) virus belonging to the family Partitiviridae and the genus Alphacryptovirus, usually associated with other viruses Mixed infections aggravate the symptoms of co-viruses on the host. In 1978, Kenten et al. first discovered the emergence of a recessive virus on broad bean (Vicia faba), which was called vetch latent virus (VCV). In 2009, Chen Jishuang discovered the latent virus of vetch in Hangzhou, Zhejiang, China, and conducted affiliation and functional analysis of the sequence. In 2020, Zhang Kun and others first discovered VCV-M infection on cowpea and broad bean grown in Jiangsu, and analyzed the genome of their isolates. However, it is generally believed that VCV infects plants without corresponding symptoms, and the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/40C12Q1/70C12Q1/6844C12N15/11C12R1/94
Inventor 徐小满崔晓艳秦嘉超陈华涛张红梅刘晓庆孙文献陈新
Owner JIANGSU ACAD OF AGRI SCI