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Preparation, detection and application of polyclonal antibody of yam virus X

A technology of polyclonal antibody and yam, which is applied in the field of genetic engineering, can solve the problems of production loss of yam, affecting the yield and quality of yam

Active Publication Date: 2014-01-29
GUANGXI UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

Viral diseases seriously affect the yield and quality of yam, causing heavy losses in the production of yam

Method used

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  • Preparation, detection and application of polyclonal antibody of yam virus X
  • Preparation, detection and application of polyclonal antibody of yam virus X
  • Preparation, detection and application of polyclonal antibody of yam virus X

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Embodiment Construction

[0047] The Escherichia coli (Escherichia coli) strains DH5α and BL21(DE3)pLysS strains used below were preserved by our laboratory, the 5' / 3'RACE kit was purchased from Roche Applied Life Sciences (Roche), and the PCR product cloning vector was purchased from Invitrogen , restriction endonuclease, PCR Mix, etc. were purchased from Thermo Company.

[0048] 1. Cloning of the full-length genome sequence of YVX

[0049] 1.1 The 3' end sequence cloning of 1YVX

[0050] According to the sequence of YVX obtained by the deep sequencing of the transcriptome, the cDNA first-strand synthetic primer N1T (5'-GACCACGCGTATCGATGTCGAC(T) complementary to the 3'poly(A) end of YVX and with oligo(dT) was designed 17 V-3', see sequence listing SEQ.ID.No.4, provided by the kit), forward primer YVX-3'-1F

[0051] (5'-TTGCTCGTCGTTCTCTGGCTGC-3', see the sequence listing SEQ.ID.No.5) and YVX-3'-2F

[0052] (5'-CCTCCTGCTAACTGGTCTGCTCTC-3', see the sequence listing SEQ.ID.No.6), reverse primer N1

[...

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Abstract

The invention discloses preparation, detection and application of a polyclonal antibody of a virus of the potato virus X potyvirus for infecting yam virus X, (yam virus X, YVX). Based on the YVX genome, prokaryotic expression is utilized by an inventor to purify the CP (coat protein) and be used for immune a rabbit to prepare the polyclonal antibody; the polyclonal antibody can specifically recognize the CP of the YVX subjected to prokaryotic expression and the CP of the YVX virus infected by yam, and can generate a specificity immune reaction. Based on the preparation, an efficient, high-flexibility and accurate IC-RT-PCR (immunocapture-reverse transcription-polymerase chain reaction), ELISA (enzyme-linked immuno sorbent assay) and Western blot method is established by the inventor, and YVX can be detected specifically from a yam plant which is infected by the YVX. According to the invention, a material basis and a technical support can be provided for the mutual interaction research of the YVX and a host and the rapid detection, parting and molecular biology research of the virus, and a solid basis is developed for the prevalence monitoring, prevention and treatment of the virus.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to the preparation, detection and application of a polyclonal antibody of Potatovirus X virus infecting Chinese yam, namely Chinese yam X virus. Background technique [0002] Yam, also known as Chinese yam and Chinese yam, is a plant of the family Dioscorea. Its roots are rich in starch, protein, vitamins, amino acids and various medicinal ingredients such as choline, saponins, mucopolysaccharides, etc., and also contain Fe, Zn, Cu, Ca, etc. A variety of minerals, depending on the species and varieties, can be used for medicine or food, with good market prospects and industrial development potential. Viral diseases have seriously affected the yield and quality of Chinese yam, and caused great losses in the production of Chinese yam. The literature shows that the viruses that infect yam under natural conditions include Dioscorea alata bacilliform virus (DaBV)...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/531
CPCG01N33/531G01N33/56983
Inventor 陈保善邹承武蒙姣荣
Owner GUANGXI UNIV
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