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Infusion protein of single chain antibodyA1 and metacide hydrolase, and its preparation method

A technology of methyl parathion and fusion protein, applied in botany equipment and methods, biochemical equipment and methods, chemical instruments and methods, etc.

Inactive Publication Date: 2008-12-31
INST OF AQUATIC LIFE ACAD SINICA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, shrimp hemolymph has strong activity to react with alkaline phosphatase and horseradish peroxidase substrates, but has no activity to react with methyl parathion hydrolase substrates

Method used

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  • Infusion protein of single chain antibodyA1 and metacide hydrolase, and its preparation method
  • Infusion protein of single chain antibodyA1 and metacide hydrolase, and its preparation method

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Embodiment Construction

[0058] The method for preparing single-chain antibody A1-methyl parathion hydrolase fusion protein with single-chain antibody A1 and methyl parathion hydrolase is carried out according to the following steps:

[0059] 1. Using the upstream primer 5'-CCCCGACTAGTATGGCCGAGGTGAAGCT-3' with the Spe I endonuclease site and the downstream primer 5'-CACCAAGCTTTTAGCGCTTGATTTCCAAC-3' with the HindIII endonuclease site, by PCR, the single-chain antibody A1 The gene was amplified from the phagemid vector pCANTAB5E-A1. The conditions of PCR are: 94°C for 5 minutes; 94°C for 45 seconds, 44°C for 1 minute, 72°C for 2 minutes, 5 cycles; 94°C for 45 seconds, 62°C for 1 minute, 72°C for 2 minutes, 25 cycles; 72°C 7 minutes; 10 minutes at 4°C. The fragments amplified by PCR were first cloned into the pGEM-T vector, and the clones connected with the A1 gene were selected by PCR and endonuclease analysis. The connected A1 gene was digested with Spe I and HindIII endonucleases, and the enzyme dig...

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Abstract

The invention discloses a single-chain antibody A1-methyl parathion hydrolase fusion protein for recognizing prawn white spot baculovirus and a preparation method thereof, and relates to an aquaculture biological virus antibody. Single-chain antibody gene and methyl parathion hydrolase gene are encoded by [-(Gly-Ser) 5 -] The oligonucleotide fragments of small peptides are connected with complementary strands to form the fusion gene of MPH-L-A1E, and then cloned into the expression vector of Escherichia coli, and the fusion protein is expressed in a secretable form. The fusion protein can be directly combined with the virus and produce a color reaction, which can save the necessary secondary antibody operation steps when using the single-chain antibody A1 that recognizes the white spot syndrome virus to detect the virus, simplifies the operation procedure, and reduces the production cost. testing cost.

Description

Technical field: [0001] The present invention relates to aquaculture biological virus antibody, more specifically relates to prawn white spot syndrome virus single chain antibody and methyl parathion hydrolase fusion protein. Background technique: [0002] Since 1993, shrimp diseases have exploded in my country, causing large-scale death of cultured prawns and heavy economic losses. Studies have shown that it is mainly caused by a new shrimp white spot syndrome virus. The virus can infect all artificially cultured prawn species in my country. It has strong infectivity, rapid onset and high mortality, which brings great harm to the prawn farming industry. At present, there is no effective treatment for shrimp virus diseases, and avoiding contact with virus sources is considered to be the most effective preventive measure, which mainly relies on early rapid diagnosis. The current diagnostic methods mainly include nucleic acid probe technology and enzyme-linked immunosorbent ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/62C12N15/13C12N15/55C12N15/09C07K19/00
Inventor 戴和平张先恩周亚凤杨薇张晓华戴玲芬张治平
Owner INST OF AQUATIC LIFE ACAD SINICA