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Information Journal Paper

Title

RECOMBINANT EXPRESSION OF LIGHT CHAIN OF BOTULINUM NEUROTOXIN TYPE-A IN E. COLI AND EVALUATION OF ITS ENZYMATIC ACTIVITY

Pages

  58-58

Abstract

 Botulinum neurotoxins, specifically type-A, are one of the most dangerous biological agents in nature that cause paralysis. These toxins have ENDOPEPTIDASE ACTIVITY which can inhibit secretion of acetylcholine neurotransmitter through neuron synapsis. This study focuses on RECOMBINANT EXPRESSION of CATALYTIC DOMAIN (light chain) of BOTULINUM NEUROTOXIN TYPE-A with high yields in order to evaluate its catalytic activity. Sequence of light chain of BOTULINUM NEUROTOXIN TYPE-A was obtained from NCBI databank.Optimizing the codon preferences for E. coli and final sequence were ordered for subcloning in pET28a (+). Vector including mentioned gene was transferred into E. Coli BL21-DE3 for expression. Protein expression conditions were optimized for solubilization of protein. Expressed protein were purified with Ni-NTA, proved with specific antibody and analyzed for its activity with HPLC technique. Best soluble expression conditions of protein are achieved in induction OD of 0.5, 0.5 mM of IPTG for 18 hours in 18o C. Soluble protein were purified with Ni-Agarode affinity chromatography and proved with western blotting and ELIZA techniques. Results represent soluble expression of light chain of BOTULINUM NEUROTOXIN TYPE-A with high yields over 98%. Analysis of HPLC data demonstrates 3 mmol min-1 mg-1 specific activity of light chain of BOTULINUM NEUROTOXIN TYPE-A.

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    APA: Copy

    MOOSAVI, SEYED JAFAR, RASHIDIANI, JAMAL, FARASAT, ALIREZA, & EBRAHIMI, FIROUZ. (2014). RECOMBINANT EXPRESSION OF LIGHT CHAIN OF BOTULINUM NEUROTOXIN TYPE-A IN E. COLI AND EVALUATION OF ITS ENZYMATIC ACTIVITY. MOLECULAR BIOLOGY RESEARCH COMMUNICATIONS, 3(SUPPL. (1)), 58-58. SID. https://sid.ir/paper/336094/en

    Vancouver: Copy

    MOOSAVI SEYED JAFAR, RASHIDIANI JAMAL, FARASAT ALIREZA, EBRAHIMI FIROUZ. RECOMBINANT EXPRESSION OF LIGHT CHAIN OF BOTULINUM NEUROTOXIN TYPE-A IN E. COLI AND EVALUATION OF ITS ENZYMATIC ACTIVITY. MOLECULAR BIOLOGY RESEARCH COMMUNICATIONS[Internet]. 2014;3(SUPPL. (1)):58-58. Available from: https://sid.ir/paper/336094/en

    IEEE: Copy

    SEYED JAFAR MOOSAVI, JAMAL RASHIDIANI, ALIREZA FARASAT, and FIROUZ EBRAHIMI, “RECOMBINANT EXPRESSION OF LIGHT CHAIN OF BOTULINUM NEUROTOXIN TYPE-A IN E. COLI AND EVALUATION OF ITS ENZYMATIC ACTIVITY,” MOLECULAR BIOLOGY RESEARCH COMMUNICATIONS, vol. 3, no. SUPPL. (1), pp. 58–58, 2014, [Online]. Available: https://sid.ir/paper/336094/en

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