Expression, Purification and Polyclonal Antibody Preparation of the Flagellin Gene fliC of Salmonella typhimurium

doi:10.7506/spkx1002-6630-201309037

FOOD SCIENCE ›› 2013, Vol. 34 ›› Issue (9): 180-184.doi: 10.7506/spkx1002-6630-201309037

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Expression, Purification and Polyclonal Antibody Preparation of the Flagellin Gene fliC of Salmonella typhimurium

CHEN Ming1,2，XU Xing-lian2，ZHOU Guang-hong2，TANG Xiao-yan1,*，YUAN Fei3，CHEN Ai-liang1

1. Key Laboratory of Agri-food Safety and Quality, Ministry of Agriculture, Institute of Quality Standards and Testing Technology
for Agri-products, Chinese Academy of Agricultural Sciences, Beijing 100081, China；2. Key Laboratory of Meat Processing and
Quality Control, Ministry of Education, College of Food Science and Technology, Nanjing Agricultural University,
Nanjing 210095, China；3. Chinese Academy of Inspection and Quarantine, Beijing 100123, China

Received:2012-05-10 Revised:2013-04-10 Online:2013-05-15 Published:2013-05-07
Contact: TANG Xiao-yan E-mail:txycaas@126.com

Abstract

Abstract:

In this study, the flagellar gene fliC of Salmonella typhimurium was amplified by PCR from the Salmonella
genome, and cloned into the prokaryotic expression vector pET28a(+). After being sequenced, the recombinant plasmid
pET28a-fliC was transformed into strain BL21(DE3)pLysS, and FliC protein was expressed by the recombinant strain after
IPTG(1 mmol/L) induction. By Ni-NTA purification, the 6×His-tagged proteins were extracted. The molecular weight of
the induced protein was about 54.6 kD as expected. And most of the expressed products were soluble. Western-blotting
analysis indicated that the expressed proteins had a specific reactivity with the monoclonal anti-His-Tag antibody. Polyclonal
antibodies were obtained via immunizing BALB/c mice with the purified protein, and the results showed that the polyclonal
antibodies had high titer and good specificity.

Key words: Slamonella typhimurium, flagellar, prokaryotic expression, purification, identification, polyclonal antibodies

CLC Number:

Q786

CHEN Ming1,2，XU Xing-lian2，ZHOU Guang-hong2，TANG Xiao-yan1,*，YUAN Fei3，CHEN Ai-liang1. Expression, Purification and Polyclonal Antibody Preparation of the Flagellin Gene fliC of Salmonella typhimurium[J]. FOOD SCIENCE, 2013, 34(9): 180-184.

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Expression, Purification and Polyclonal Antibody Preparation of the Flagellin Gene fliC of Salmonella typhimurium

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