Vitellogenin peptide fragment tfVWD used for binding to tetrodotoxin, nucleotide sequence of vitellogenin peptide fragment tfVWD, polyclonal antibody of vitellogenin peptide fragment tfVWD and preparation method for vitellogenin peptide fragment tfVWD

Abstract

The present invention discloses a vitellogenin peptide fragment tfVWD used for binding to tetrodotoxin. The vitellogenin peptide fragment tfVWD comprises a polypeptide having an amino acid sequence asshown in SEQ ID NO: 1, or an active fragment of the polypeptide. The invention further discloses a nucleotide sequence encoding the vitellogenin peptide fragment tfVWD used for binding to the tetrodotoxin; as shown in SEQ ID NO:2, by taking the vitellogenin peptide fragment tfVWD as an antigen, and mice are immunized to prepare antiserum to obtain a polyclonal antibody. The invention further discloses a preparation method for the vitellogenin peptide fragment tfVWD, and the preparation method comprises the following steps: S1, the nucleotide sequence encoding the vitellogenin peptide fragmenttfVWD is obtained by cloning, and the nucleotide sequence is cloned to a recombinant expression vector; S2, induced expression and condition optimization are performed on the recombinant expression vector to obtain an expression product; and S3, the expression product is filtered, and is eluted through an affinity chromatography column, and elution peaks are collected to obtain recombinant protein. The vitellogenin peptide fragment tfVWD and the nucleotide sequence and polyclonal antibody thereof obtained by the invention have a good binding property with the tetrodotoxin (TTX), and lay a foundation for developing detoxification therapeutic drugs for the tetrodotoxin.

Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a vitellogenin peptide (tfVWD), its encoded nucleotide sequence and its polyclonal antibody. The invention also relates to a preparation method of the vitellogenin peptide tfVWD. Background technique

[0002] Tetrodotoxin (TTX) is a small molecule alkaloid neurotoxin with the molecular formula C 11 h 17 o 8 N 3 , with a molecular weight of 319. It is an amino perhydroquinazoline compound and is one of the most toxic neurotoxins found in nature. Tetrodotoxin is a typical Na + Channel blocker, can interact with Na on the surface of muscle nerve cell membrane + Channel receptor specific binding, inhibiting Na in nerve cells + influx and block excitatory transmission between nerve and muscle.

[0003] Poisoning incidents caused by tetrodotoxin often occur in China, Japan and some countries in Southeast Asia, mainly due to eating poisonous pufferfish by mistake. Scholars at home...

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