مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

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Information Journal Paper

Title

PRODUCTION AND CHARACTERIZATION OF RECOMBINANT LIGHT CHAIN AND CARBOXYTERMINAL HEAVY CHAIN FRAGMENTS OF TETANUS TOXIN

Pages

  220-226

Abstract

 Background: LIGHT CHAIN (LC) and heavy chain carboxyterminal subdomain (HCC) fragments are the most important parts of tetanus neurotoxin (TeNT) which play key roles in toxicity and binding of TeNT, respectively. In the present study, these two fragments were cloned and expressed in a prokaryotic system and their identity was confirmed using anti-TeNT specific polyclonal and monoclonal antibodies.Methods: LC and HCC gene segments were amplified from Clostridium tetani genomic DNA by PCR, cloned into pET28b (+) cloning vector and transformed in Escherichia coli (E. coli) BL21 (DE3) expression host. Recombinant proteins were then purified through His-tag using Nickel-based chromatography and characterized by SDS-PAGE, Western blotting and ELISA techniques.Results: Recombinant LIGHT CHAIN and HCC fragments were successfully cloned and expressed in (E. coli) BL21 (DE3). Optimization of the induction protocol resulted in production of high levels of HCC (~35% of total bacterial protein) and to lesser extends of LC (~5%). Reactivity of the His-tag purified proteins with specific polyclonal and monoclonal antibodies confirmed their renatured structure and identity.Conclusion: Our results indicate successful cloning and production of recombinant LC and HCC fragments of TeNT. These two recombinant proteins are potentially useful tools for screening and monitoring of anti-TeNT antibody response and vaccine production.

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    Cite

    APA: Copy

    YOUSEFI, MEHDI, KHOSRAVI EGHBAL, ROYA, HEMMATI, AZAM, & SHOKRI, FAZEL. (2013). PRODUCTION AND CHARACTERIZATION OF RECOMBINANT LIGHT CHAIN AND CARBOXYTERMINAL HEAVY CHAIN FRAGMENTS OF TETANUS TOXIN. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), 5(4 (18)), 220-226. SID. https://sid.ir/paper/313817/en

    Vancouver: Copy

    YOUSEFI MEHDI, KHOSRAVI EGHBAL ROYA, HEMMATI AZAM, SHOKRI FAZEL. PRODUCTION AND CHARACTERIZATION OF RECOMBINANT LIGHT CHAIN AND CARBOXYTERMINAL HEAVY CHAIN FRAGMENTS OF TETANUS TOXIN. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)[Internet]. 2013;5(4 (18)):220-226. Available from: https://sid.ir/paper/313817/en

    IEEE: Copy

    MEHDI YOUSEFI, ROYA KHOSRAVI EGHBAL, AZAM HEMMATI, and FAZEL SHOKRI, “PRODUCTION AND CHARACTERIZATION OF RECOMBINANT LIGHT CHAIN AND CARBOXYTERMINAL HEAVY CHAIN FRAGMENTS OF TETANUS TOXIN,” AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), vol. 5, no. 4 (18), pp. 220–226, 2013, [Online]. Available: https://sid.ir/paper/313817/en

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