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Information Journal Paper

Title

CLONING AND EXPRESSION OF S1 SUBUNIT OF PERTUSSIS TOXIN IN ESCHERICHIA COLI

Pages

  19-24

Abstract

 BORDETELLA PERTUSSIS is a gram negative bacterium that causes respiratory tract infection in human (whooping cough). PERTUSSIS TOXIN (PT) is the main component of current acellular pertussis vaccine and the S1 (subunit1) is the main immunogenic part of it. Thus, S1 has been the target of many studies as a potent candidate of acellular vaccine against BORDETELLA PERTUSSIS, lacking the side effects of whole cell based ones. S1 gene was amplified and inserted in three expression vectors including pET-14b, pET-22b (+) and pAED4. The possibility and level of expression of these constructs were investigated in BL21 (DE3) strain of ESCHERICHIA COLI (E.coli) as expression host. The highest expression was in pET-22b (+) -S1. Best expression achieved 6 hr post induction with 0.2 mM IPTG in LB broth containing ampicillin, at 30oC with shaking (250 rpm). Recombinant S1 protein was observed in two distinct separated proteins with 28 and 31 kDa estimated molecular weight. In spite of toxicity of PT and S1 in the E.coli, considerable amount of S1 was expressed in E.coli. Two rS1 bands were detected on SDS-PAGE. Both were confirmed as S1 in western blot with specific monoclonal and polyclonal antibodies against PERTUSSIS TOXIN. Appearance of two distinct bands could be the result of leader peptidase activity or nonspecific peptidase from E.coli on recombinant S1. As the recombinant S1 is a suitable antigen for studies as a candidate acellular vaccine or development of ELISA for detection of BORDETELLA PERTUSSIS, further studies are underway.

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    APA: Copy

    KHAFRI, ABOLFAZL, AGHAIYPOUR, KHOSROW, NAJAR PEERAYEH, SHAHIN, & GHORBANI, REIHANEH. (2011). CLONING AND EXPRESSION OF S1 SUBUNIT OF PERTUSSIS TOXIN IN ESCHERICHIA COLI. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), 3(1 (8)), 19-24. SID. https://sid.ir/paper/313749/en

    Vancouver: Copy

    KHAFRI ABOLFAZL, AGHAIYPOUR KHOSROW, NAJAR PEERAYEH SHAHIN, GHORBANI REIHANEH. CLONING AND EXPRESSION OF S1 SUBUNIT OF PERTUSSIS TOXIN IN ESCHERICHIA COLI. AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB)[Internet]. 2011;3(1 (8)):19-24. Available from: https://sid.ir/paper/313749/en

    IEEE: Copy

    ABOLFAZL KHAFRI, KHOSROW AGHAIYPOUR, SHAHIN NAJAR PEERAYEH, and REIHANEH GHORBANI, “CLONING AND EXPRESSION OF S1 SUBUNIT OF PERTUSSIS TOXIN IN ESCHERICHIA COLI,” AVICENNA JOURNAL OF MEDICAL BIOTECHNOLOGY (AJMB), vol. 3, no. 1 (8), pp. 19–24, 2011, [Online]. Available: https://sid.ir/paper/313749/en

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