49 results about "Scorpion toxin" patented technology

The invention discloses a purifying and refolding method for a scorpion toxin protein inclusion body. The method mainly comprises: expressing a scorpion toxin protein by virtue of recombinant escherichia coli, wherein the terminal C of the scorpion toxin protein contains His-tag; then opening a disulfide bond in the scorpion toxin protein by using a denaturation buffer solution; purifying the denatured scorpion toxin protein by virtue of a histidine affinity column; and then performing renaturation on the scorpion toxin protein by using a renaturation buffer solution, wherein the renaturation buffer solution comprises 50-200 mmol / L Na2HPO4, 10-100 mmol / L Tris, 0.1-1 mol / L L-Arg, 1-5 mmol / L EDTA, 0.1-5 mmol / L GSH, 0.05-0.5 mmol / L GSSG, 5-20% (v / v) of glycerinum and 0.01-5% (v / v) of tritonX-100, and the pH is 7-9. The scorpion toxin protein is prepared by the method disclosed by the invention, and the method has the advantages of being simple to operate and good in renaturation effects.

Provided are optimized polynucleotide sequences encoding a scorpion Androctonus australis neurotoxin AaIT, which are suitable for being transcribed and expressed in fungi. Also provided are the fungi capable of expressing AaIT, such as Metarhizium anisopliae, which have improved insecticidal efficiency and can be used for insect control in effect.

The invention discloses a scorpion toxin active polypeptide HsTx1 capable of improving cerebral ischemia and hypoxia, and a preparation method and application thereof. The amino acid sequence of the scorpion toxin active polypeptide HsTx1 with improving cerebral ischemia and hypoxia is AGKKERAGSRRTKIVMLKCIREHGH. The tropical rainforest scorpion toxin polypeptide HsTx1 purified by the present invention shows strong anti-cerebral ischemia and hypoxia (MCAO) activities and improves motor dysfunction in the cerebral ischemia and hypoxia model (MCAO) experiment, indicating that the "scorpion toxin of the present invention can improve cerebral ischemia and hypoxia." Hypoxic active polypeptide HsTx1" has great medical application prospects. The scorpion toxin active polypeptide HsTx1 with improving cerebral ischemia and hypoxia described in the present invention has the beneficial characteristics of strong activity of effectively improving cerebral ischemia hypoxia and motor dysfunction.

The invention discloses a method for producing recombinant scorpion toxin protein by adopting a silkworm as a parasitifer, which mainly comprises the following steps: adopting a scorpion cDNA as a template, adopting 5'-ACGTAGATCTATGGATGGATATATAAG-3' containing BglII enzyme cutting site as a forward primer and adopting 5'-ACGTAGATCT TTACTTTTTTCCAC-3' containing the BglII enzyme cutting site as a reverse primer to carry out PCR gene amplification reaction in 32 cycles to obtain a scorpion toxin gene; digesting pCR2.1-scorpion toxin by restriction endonuclease BglII to obtain a scorpion toxin gene fragment; cloning the scorpion toxin gene fragment in a baculovirus transfer vector pAcGP67b to obtain a recombinant pAcGP67b-scorpion toxin gene, carrying out homologous recombination with baculovirus BmNPV DNA of a silkworm in an insect cell by cotransfection to produce a recombinant virus; expressing in a silkworm body and purifying the recombinant scorpion toxin. Most of the recombinant scorpion toxin protein expressed by a silkworm larva is retained in a fat body of the silkworm; the scorpion toxin is purified from the fat body by an affinity chromatography; and the recombinant scorpion toxin has obvious biological toxicity, is suitable for the requirement of a biopharmaceutical industrialization level and has wide application prospect in the field of future medical and clinic treatment.

The invention belongs to the field of medical biology, and particularly relates to a production method for recombining scorpion toxin protein AGAP. The method comprises the following steps: firstly, obtaining SUMO-AGAP fusion genes with His6 labels by using overlapping PCR technology; secondly, constructing a recombinant expression vector containing the SUMO-AGAP fusion genes; thirdly, transferring the constructed expression vector to a host cell to express; and finally, cracking the cell by using a physical method or a chemical agent, extracting recombinant fusion protein and His purified fusion protein to obtain pure SUMO-AGAP fusion protein, carrying out enzyme cutting reaction, releasing AGAP, and further purifying the fusion protein to obtain the purified AGAP protein. The method greatly improves the yield, saves the cost, and is simple and convenient to operate; and the AGAP produced by the method has broad biologic activity, can restrain the proliferation of various tumor cells, and simultaneously has analgesia effect.