Primers, kit and method for identifying potato M viruses

An identification method, potato technology, applied in biochemical equipment and methods, microbe determination/inspection, DNA/RNA fragments, etc., can solve problems such as unclear functions

Inactive Publication Date: 2017-10-03
陈定虎
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

ORFI of PVM RNA encodes a 223kDa replicase; ORF2, ORF3, and ORF4 encode a 25kDa, 12kDa, and 7kDa protein respectively, forming a three-gene cassette that participates in the intercellular movement of the virus; ORF5 encodes a 34kDa coat protein; overlapping ORF6 Encodes a cysteine-rich protein of 11-16kDa. The function of this product is unclear, but from the ability to bind nucleic acids, it may help aphids spread or be involved in host gene transcription and viral RNA replication[1]

Method used

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  • Primers, kit and method for identifying potato M viruses
  • Primers, kit and method for identifying potato M viruses
  • Primers, kit and method for identifying potato M viruses

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0076] A kit for identifying potato M virus RT-LAMP, the kit includes:

[0077]

[0078] Among them, the primer sequence:

[0079] PVM-F3:5'-TTCAGGCCGTGCTGTTCT-3'

[0080] PVM-B3:5'-CAGCATGTGGTTCCAAGTCA-3'

[0081] PVM-FIP:5'-GCACCCCTAGGCCACTCAAA-GGATGCAAGCAGCTCAGT-3'

[0082] PVM-BIP: 5'-GGACGCTGTGCTTGCTGTGA-CAGGCGCATACAACCTACA-3'.

Embodiment 2

[0084] A kit for identifying potato M virus RT-LAMP, the kit includes:

[0085]

[0086] Among them, the primer sequence:

[0087] PVM-F3:5'-TTCAGGCCGTGCTGTTCT-3'

[0088] PVM-B3:5'-CAGCATGTGGTTCCAAGTCA-3'

[0089] PVM-FIP:5'-GCACCCCTAGGCCACTCAAA-GGATGCAAGCAGCTCAGT-3'

[0090] PVM-BIP: 5'-GGACGCTGTGCTTGCTGTGA-CAGGCGCATACAACCTACA-3'.

[0091] The 2× reaction solution buffer described in the present invention is made up of following components:

[0092]

[0093]

[0094] The enzyme solution is a mixture of Bst DNA polymerase and AMV reverse transcriptase.

[0095] The fluorescent visual detection reagent contains calcein fluorescent dye.

Embodiment 3

[0097] A kit for identifying potato M virus RT-LAMP, the kit includes:

[0098]

[0099] Among them, the primer sequence:

[0100] PVM-F3:5'-TTCAGGCCGTGCTGTTCT-3'

[0101] PVM-B3:5'-CAGCATGTGGTTCCAAGTCA-3'

[0102] PVM-FIP:5'-GCACCCCTAGGCCACTCAAA-GGATGCAAGCAGCTCAGT-3'

[0103] PVM-BIP: 5'-GGACGCTGTGCTTGCTGTGA-CAGGCGCATACAACCTACA-3'.

[0104] The 2× reaction solution buffer described in the present invention is made up of following components:

[0105]

[0106]

[0107] The enzyme solution is a mixture of Bst DNA polymerase and AMV reverse transcriptase.

[0108] The fluorescent visual detection reagent contains calcein fluorescent dye.

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Abstract

The invention discloses primers, kit and method for identifying potato M viruses RT-LAMP. The primers comprise PVM-F3: 5'-TTCAGGCCGTGCTGTTCT-3', PVM-B3: 5'-CAGCATGTGGTTCCAAGTCA-3', PVM-FIP: 5'-GCACCCCTAGGCCACTCAAA-GGATGCAAGCAGCTCAGT-3' and PVM-BIP: 5'-GGACGCTGTGCTTGCTGTGA-CAGGCGCATACAACCTACA-3'. In order to solve the problems of the prior art, the invention provides a primer method, kit and a method for rapid, simple, efficient, high accuracy, high specificity, high sensitivity identification of potato M viruses RT-LAMP.

Description

technical field [0001] The invention relates to a primer, a kit and a detection method for identifying potato M virus RT-LAMP, and belongs to the field of biotechnology. Background technique [0002] Potato virus M, PVM was found in the United States, France, the United Kingdom, Germany, the Netherlands and other countries. At present, PVM occurs all over the world [1]. PVM is a member of Carlavirus genus Carlavirus. The size of the virion is 650nmX12nm, and there is an incompletely closed viral capsid. The lethal temperature is 65-71°C, the in vitro preservation period is 5-7 days, and the dilution limit is 10 2 -10 3 . The virus could be detected in any part of the host plant, and the virus mainly existed in the cytoplasm. Amorphous X-body-shaped inclusion bodies were found in the cytoplasm of susceptible plant cells. The virus genome is a single-stranded positive-sense RNA with a length of 8535bp. There is a Poly(A) at the 3' end and a methylated cap structure at th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/70C12Q1/68C12N15/11
CPCC12Q1/6844C12Q1/701C12Q2531/119C12Q2521/107
Inventor 刘洪义陈定虎陈文聪刘运张静苏斐
Owner 陈定虎
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