Cloning of fliC Gene From Salmonella typhimurium in the Expression Vector pVAX1 and Evaluation of its Expression in Eukaryotic Cells

Jundishapur Journal of Microbiology
Reza TaherkhaniAli Reza Samarbafzadeh

Abstract

Flagellin is the main structural protein of the flagella of many pathogens including Salmonella typhimurium. It is a potent trigger of innate immune responses that enhance adaptive immune responses to a variety of protein antigens. Flagellin has intrinsic adjuvant activity mediated through toll-like receptor (TLR) 5 and is an attractive candidate for highly effective vaccine adjuvant conferring enhanced antibody and cellular immune responses to proteins or peptides. In the present study, we cloned the fliC gene from S. enterica typhimurium in eukaryote vector pVAX1 and evaluated its expression in eukaryotic cells. The main aim of the present study was to construct a DNA vaccine expressing fliC as an adjuvant. The fliC gene of S. typhimurium (ATCC 14028) was amplified by PCR with specific primers and cloned into the pPrime cloning vector and successfully subcloned into expression vector pVAX1. The recombinant plasmid pVAX-fliC was finally expressed in eukaryotic cells. Cloning and subcloning of the fliC gene were confirmed by colony PCR, restriction enzymes digestion and DNA sequencing of the recombinant plasmids pPrime-fliC and pVAX-fliC. The expression of flagellin protein in eukaryotic cells was approved by immunofluorescence a...Continue Reading

References

Aug 8, 2001·The Journal of Immunology : Official Journal of the American Association of Immunologists·A T GewirtzJ L Madara
Feb 28, 2002·Annual Review of Immunology·Arthur M Krieg
Mar 23, 2002·Biochimica Et Biophysica Acta·Tsuneyasu Kaisho, Shizuo Akira
Jan 14, 2003·Annual Review of Immunology·Kiyoshi TakedaShizuo Akira
Nov 26, 2003·Nature Immunology·Jean-Marc Reichhart
Apr 23, 2004·Infection and Immunity·Camilo CuadrosJoseph Lustgarten
Dec 15, 2004·Microbes and Infection·Chandrashekhar Pasare, Ruslan Medzhitov
Aug 27, 2005·Immunologic Research·Anna N Honko, Steven B Mizel
Sep 9, 2005·The Journal of Immunology : Official Journal of the American Association of Immunologists·Steven E ApplequistHans-Gustaf Ljunggren
Dec 2, 2005·Current Opinion in Gastroenterology·Andrew T Gewirtz
Feb 25, 2006·Cell·Shizuo AkiraOsamu Takeuchi
Feb 23, 2007·Immunology·Danielle AwDonald B Palmer
May 2, 2007·The Journal of Infectious Diseases·William F McDonaldT J Powell
Nov 24, 2007·Expert Review of Vaccines·Tamar Ben-Yedidia, Ruth Arnon
Mar 28, 2008·Mechanisms of Ageing and Development·John T BatesSteven B Mizel
Jun 5, 2008·The Journal of Immunology : Official Journal of the American Association of Immunologists·Matam Vijay-KumarAndrew T Gewirtz
Oct 30, 2010·Immunity·Robert L CoffmanRobert A Seder
Nov 5, 2010·The Journal of Immunology : Official Journal of the American Association of Immunologists·Steven B Mizel, John T Bates
Jan 10, 2012·Veterinary Microbiology·Hso-Chi ChaungYi-Yang Lien

❮ Previous
Next ❯

Citations


❮ Previous
Next ❯

Datasets Mentioned

BETA
2059063-2060550

Methods Mentioned

BETA
PCR
electrophoresis

Related Concepts

Related Feeds

Avian Influenza: Innate Immune Adjuvant (ASM)

Adjuvants systems that are added to vaccines against avian influenza have be explored to enhance the innate immune system response against the virus. Here is the latest research on avian influenza and the innate immune adjuvant.

Avian Influenza: Innate Immune Adjuvant

Adjuvants systems that are added to vaccines against avian influenza have be explored to enhance the innate immune system response against the virus. Here is the latest research on avian influenza and the innate immune adjuvant.

Cancer Vaccines

Cancer vaccines are vaccines that either treat existing cancer or prevent development of a cancer.