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Nano antibody for specifically recognizing vibrio parahaemolyticus, recombinant vector, host cell and application thereof

A hemolytic vibrio and nanobody technology, which is applied in the field of microbial detection to achieve the effects of good specificity, high expression yield and strong binding ability

Active Publication Date: 2021-10-01
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, among the existing nanobodies, there is no nanobody against Vibrio parahaemolyticus, and the detection technology of Vibrio parahaemolyticus using this nanobody has been proposed

Method used

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  • Nano antibody for specifically recognizing vibrio parahaemolyticus, recombinant vector, host cell and application thereof
  • Nano antibody for specifically recognizing vibrio parahaemolyticus, recombinant vector, host cell and application thereof
  • Nano antibody for specifically recognizing vibrio parahaemolyticus, recombinant vector, host cell and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0031] 1. Construction of anti-Vibrio parahaemolyticus phage display nanobody library

[0032] 1.1 Immunity of camels:

[0033] Select a healthy camel that has not been immunized with any antigen, and mix an equal volume of emulsified mixture of inactivated Vibrio parahaemolyticus and Freund's complete adjuvant in 10 8 Bactrian camels were first immunized at a concentration of CFU / mL, and the antigen was emulsified with incomplete Freund's adjuvant for subsequent immunizations, and boosted once every two weeks for a total of 6 immunizations. One week after each immunization, the camel blood was collected to detect the antibody titer in the serum.

[0034] 1.2 Extraction of total RNA from blood: After the fifth immunization, the peripheral blood of camels was collected, and total RNA was extracted according to the operation steps of the RNA extraction kit.

[0035] 1.3 Reverse transcription to obtain cDNA:

[0036] The obtained total RNA is used as a template, oligo(dT) 15 ...

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Abstract

The invention provides a nano antibody for specifically recognizing vibrio parahaemolyticus, which comprises a coding gene of the nano antibody, and also provides a vector and a host cell containing a nucleotide sequence for coding the nano antibody. The amino acid sequence of the nano antibody is shown as SEQ ID NO: 1, and the nano antibody has good specificity and binding capacity to vibrio parahaemolyticus. The nano antibody for specifically recognizing the vibrio parahaemolyticus, provided by the invention, is good in specificity, strong in stability and high in expression yield, is not combined with other non-vibrio parahaemolyticus, and can specifically recognize the vibrio parahaemolyticus.

Description

technical field [0001] The invention belongs to the field of microorganism detection. Specifically, the present invention relates to nanobodies, recombinant vectors, host cells and applications thereof that specifically recognize Vibrio parahaemolyticus. Background technique [0002] Vibrio parahaemolyticus is a foodborne pathogen that clinically causes abdominal pain, vomiting, and diarrhea. Eating food containing this bacteria can cause food poisoning, also known as halophilic bacteria food poisoning. Vibrio parahaemolyticus is a Gram-negative bacillus with arc-shaped, rod-shaped, filamentous and other shapes, without tooth spores. It is also a marine bacterium, mainly derived from seafood such as fish, shrimp, crab, shellfish and seaweed. With the increasing market demand for seafood, in order to ensure the safety of all food processing links from fresh food to the dining table, and its low infectious dose to seafood, a rapid and sensitive detection method is essential...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/12G01N33/535G01N33/543
CPCC07K16/1239G01N33/54306G01N33/535G01N33/54313C07K2317/569C07K2317/565C07K2317/567C07K2317/52C07K2317/92G01N2333/28
Inventor 王妍入王鹏廖星蕊张瑶魏娟王建龙马敏
Owner NORTHWEST A & F UNIV
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