Nanobody, recombinant vector, host cell and application thereof for specific recognition of Vibrio parahaemolyticus

A hemolytic vibrio and nanobody technology, which is applied in the field of microbial detection to achieve the effects of accurate detection results, high thermal stability and wide application

Active Publication Date: 2022-05-27
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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  • Nanobody, recombinant vector, host cell and application thereof for specific recognition of Vibrio parahaemolyticus
  • Nanobody, recombinant vector, host cell and application thereof for specific recognition of Vibrio parahaemolyticus
  • Nanobody, recombinant vector, host cell and application thereof for specific recognition of Vibrio parahaemolyticus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

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

[0032] 1.1 Immunity of camels:

[0033] A healthy camel that has not been immunized with any antigen was selected and an equal volume of an emulsified mixture of inactivated Vibrio parahaemolyticus and Freund's complete adjuvant was prepared at a concentration of 10 8 Bactrian camels were initially immunized at the concentration of CFU / mL, and the antigens were emulsified with incomplete Freund's adjuvant for subsequent immunization. One week after each immunization, camel blood was collected to detect antibody titers in serum.

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

[0035] 1.3 Reverse transcription to obtain cDNA:

[0036] To obtain the total RNA as a template, oligo(dT) 15 Perform reverse transcription for primers, ...

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Abstract

The present invention provides a nanobody that specifically recognizes Vibrio parahaemolyticus, including the coding gene of the nanobody, and also provides a vector and a host cell containing a nucleotide sequence encoding the nanobody, and its amino acid sequence is as SEQ ID NO: As shown in 1, the nanobody has good specificity and binding ability to Vibrio parahaemolyticus. The nanobody specifically recognizing Vibrio parahaemolyticus provided by the invention has good specificity, strong stability, high expression yield, does not bind to other non-parahaemolytic Vibrio, and can specifically recognize 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 their applications that specifically recognize Vibrio parahaemolyticus. Background technique [0002] Vibrio parahaemolyticus is a food-borne pathogen that can clinically cause abdominal pain, vomiting, and diarrhea. Eating food containing this bacteria can cause food poisoning, also known as halophilic food poisoning. Vibrio parahaemolyticus is a gram-negative bacillus with arc-shaped, rod-shaped, filamentous and other shapes, without dental spores. It is also a marine bacteria, 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 table, and its low infectious dose to seafood, a rapid and sensitive detection method is essential to ensure the saf...

Claims

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Application Information

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Patent Type & Authority Patents(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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