CLONING, EXPRESSION AND PURIFICATION OF THE FACTOR H BINDING PROTEIN AND ITS INTERACTION WITH FACTOR H

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YARIAN FATEMEH; BANDEHPOUR MOJGAN; SEYED NEGAR; KAZEMI BAHRAM

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Journal Paper

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Journal: Iranian Journal of Microbiology Year:2016 | Volume:8 | Issue:1 Page(s): 29-35

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Title

CLONING, EXPRESSION AND PURIFICATION OF THE FACTOR H BINDING PROTEIN AND ITS INTERACTION WITH FACTOR H

Author(s)

YARIAN FATEMEH | BANDEHPOUR MOJGAN | SEYED NEGAR | KAZEMI BAHRAM | Issue Writer Certificate

Keywords

N. MENINGITIDIS

FHBP

CLONING

FH PROTEIN

FAR-WESTERN BLOTTING

Abstract

Background and Objective: Neisseria meningitidis is a leading cause of meningitis and sepsis worldwide. The factor H binding protein (fHBP) is a key virulence factor of Neisseria meningitidis that is able to selectively bind to human factor H, the key regulator of the alternative complement pathway, which it has important implications for meningococcal pathogenesis and vaccine design. The aims of present research were CLONING, expression, purification of FHBP and confirmation of the interaction between serum factor H (fH) and produced factor H binding protein. Materials and Methods: A 820 base pairs FHBP gene fragment was amplified by PCR and cloned into expression vector pET28a (+) in Bam HI and SalI restriction enzymes sites. Recombinant DNA was expressed in BL21 (DE3) cell. FHBP protein was purified by Ni-NTA agarose resin. Coupling of recombinant protein into CNBr activated Sepharose 4B resin was carried out for application in serum FH PROTEIN purification. (fH-fHBP) interaction was confirmed by SDS-PAGE and FAR-WESTERN BLOTTING.Results and Conclusions: SDS-PAGE results showed a 35 kDa protein band. 150 kDa FH PROTEIN was purified by designed Sepharose 4B resin. FAR-WESTERN BLOTTING confirmed (fH-fHBP) interaction and proper folding of factor H binding protein.

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APA: Copy

YARIAN, FATEMEH, BANDEHPOUR, MOJGAN, SEYED, NEGAR, & KAZEMI, BAHRAM. (2016). CLONING, EXPRESSION AND PURIFICATION OF THE FACTOR H BINDING PROTEIN AND ITS INTERACTION WITH FACTOR H. IRANIAN JOURNAL OF MICROBIOLOGY, 8(1), 29-35. SID. https://sid.ir/paper/315395/en

Vancouver: Copy

YARIAN FATEMEH, BANDEHPOUR MOJGAN, SEYED NEGAR, KAZEMI BAHRAM. CLONING, EXPRESSION AND PURIFICATION OF THE FACTOR H BINDING PROTEIN AND ITS INTERACTION WITH FACTOR H. IRANIAN JOURNAL OF MICROBIOLOGY[Internet]. 2016;8(1):29-35. Available from: https://sid.ir/paper/315395/en

IEEE: Copy

FATEMEH YARIAN, MOJGAN BANDEHPOUR, NEGAR SEYED, and BAHRAM KAZEMI, “CLONING, EXPRESSION AND PURIFICATION OF THE FACTOR H BINDING PROTEIN AND ITS INTERACTION WITH FACTOR H,” IRANIAN JOURNAL OF MICROBIOLOGY, vol. 8, no. 1, pp. 29–35, 2016, [Online]. Available: https://sid.ir/paper/315395/en

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