Volume 21 Issue 1
Jan.  2000
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QIAN You-cun, FAN Chun-yang, HU Tai-shan, YANG Yun-gui, YANG Sheng-li, GONG Yi. Molecular Cloning and Functional Expression of α-Bungarotoxin (V31) From Chinese Continental Banded Krait. Zoological Research, 2000, 21(1): 41-47.
Citation: QIAN You-cun, FAN Chun-yang, HU Tai-shan, YANG Yun-gui, YANG Sheng-li, GONG Yi. Molecular Cloning and Functional Expression of α-Bungarotoxin (V31) From Chinese Continental Banded Krait. Zoological Research, 2000, 21(1): 41-47.

Molecular Cloning and Functional Expression of α-Bungarotoxin (V31) From Chinese Continental Banded Krait

  • Received Date: 1900-01-01
  • Rev Recd Date: 1900-01-01
  • Publish Date: 2000-02-22
  • The cDNA encoding a variant of α-bungarotoxin was cloned from the venom glands of Bungarus multicinctus by RT-PCR.The deduced protein precursor contained a 21 amino acid signal peptide and a following 74 amino acid mature protein.The signal peptide is very similar to those of short chain neurotoxins,κ-neurotoxins and cardiotoxins.The amino acid sequence of the mature protein is identical to α-bungarotoxin (V31),a minor variant of α-bungarotoxin identified by protein sequencing technique.Furthermore,the cDNA encoding the deletion precursor of α-bungarotoxin was also cloned.By use of pMAL-p2,the variant was overexpressed in E.coli as a soluble fusion protein and purified by sepharose 6B-amylose affinity chromatography,which was confirmed by western blotting with the antisera against α-bungarotoxin.The recombinant variant was achieved after digestion by factor X[a].It displayed about 1/6 in vivo toxicity of natural α-bungarotoxin.The successful cloning and functional expression of α-bungarotoxin provided a basis for the future study of structure-function of long neurotoxins.
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    沈阳化工大学材料科学与工程学院 沈阳 110142

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Molecular Cloning and Functional Expression of α-Bungarotoxin (V31) From Chinese Continental Banded Krait

Abstract: The cDNA encoding a variant of α-bungarotoxin was cloned from the venom glands of Bungarus multicinctus by RT-PCR.The deduced protein precursor contained a 21 amino acid signal peptide and a following 74 amino acid mature protein.The signal peptide is very similar to those of short chain neurotoxins,κ-neurotoxins and cardiotoxins.The amino acid sequence of the mature protein is identical to α-bungarotoxin (V31),a minor variant of α-bungarotoxin identified by protein sequencing technique.Furthermore,the cDNA encoding the deletion precursor of α-bungarotoxin was also cloned.By use of pMAL-p2,the variant was overexpressed in E.coli as a soluble fusion protein and purified by sepharose 6B-amylose affinity chromatography,which was confirmed by western blotting with the antisera against α-bungarotoxin.The recombinant variant was achieved after digestion by factor X[a].It displayed about 1/6 in vivo toxicity of natural α-bungarotoxin.The successful cloning and functional expression of α-bungarotoxin provided a basis for the future study of structure-function of long neurotoxins.

QIAN You-cun, FAN Chun-yang, HU Tai-shan, YANG Yun-gui, YANG Sheng-li, GONG Yi. Molecular Cloning and Functional Expression of α-Bungarotoxin (V31) From Chinese Continental Banded Krait. Zoological Research, 2000, 21(1): 41-47.
Citation: QIAN You-cun, FAN Chun-yang, HU Tai-shan, YANG Yun-gui, YANG Sheng-li, GONG Yi. Molecular Cloning and Functional Expression of α-Bungarotoxin (V31) From Chinese Continental Banded Krait. Zoological Research, 2000, 21(1): 41-47.

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