Expression and assembly of chimeric flagellins in Escherichia coli BL21(DE3) and Salmonella.

Maozhi HU; Zhiming Pan; Yun Yang; Chuang Meng; Lei Zhang; Shizhong Geng; Xin'an Jiao

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Expression and assembly of chimeric flagellins in Escherichia coli BL21(DE3) and Salmonella.

Author: Maozhi HU ¹ ; Zhiming PAN ¹ ; Yun YANG ¹ ; Chuang MENG ¹ ; Lei ZHANG ¹ ; Shizhong GENG ¹ ; Xin'an JIAO ¹
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1. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, Jiangsu, China.
Publication Type:Journal Article
Keywords: assembly; expression; flagellin; interleukin 1β
From: Chinese Journal of Biotechnology 2017;33(8):1335-1342
CountryChina
Language:Chinese
Abstract: Flagellin can be expressed in monomeric or polymeric form based on assembly. The difference of these two forms of flagellin is less studied. In this experiment, recombinant plasmid pET-fliC/M2e2 was transferred into Escherichia coli BL21(DE3) and Salmonella SL5928 to express chimeric flagellin, mfliC/M and pfliC/M, respectively, and then their assembly characteristics were analyzed. Sodium dodecyl sulfate polyacrylamide gel electrophoresis results indicated that the two recombinant bacteria could successfully express chimeric flagellin. The transmission electronic microscope observation showed that no flagella were found on the surface of recombinant E. coli, whereas it was found for recombinant Salmonella. After purification, distinct circular dichroism spectra between them were found and pfliC/M showed the similar structure as wild-type flagellin, but not for mfliC/M. The dynamic light scattering assay also indicated that the polymerization of mfliC/M was much lower than that for pfliC/M. Three hours after transfection into mouse peritoneal macrophages, both could induce interleukin 1β secretion, but mfliC/M is stronger than pfliC/M. These data will be helpful for the selection of expression form of flagellin.