[Cloning, expression and characterization analysis of the arginine deiminase of Streptococcus suis of China isolates].
PCR analysis demonstrated the presence of the ad gene in all 29 S. suis strains tested, but none of the seven S. equi subsp. zooepidemicus strains. The fragment of ad gene of virulent isolate SS2-HA9801 was later cloned into pBAD/Myc-HisC vector via restriction endonuclease and then transformed into host strain TOP10. A recombinant protein of 47000Da was highly expressed after induced by L-ararose and purified by Ni-nitrilotriacetic acid affinity chromatography. Western blotting demonstrated that the recombinant protein can reacted to the polyclonal antibody raised against whole-cell protein of SS2-HA9801, which suggested that it possessed some immunogenicity and may be important for further research. Enzymatic assay revealed that the optimum temperature for its activity is 37 degrees C and pH is 6.5. Studies with class-specific inhibitors supported the assignment of a sulfhydryl enzyme with some metallo class characteristics.
Key Laboratory of Animal Disease Diagnostic and Immunology, Ministry of Agriculture University, Nanjing 210095, China. jqzh03@yahoo.com.cn
Jin-Qiu
JQ
Zhang
Cheng-Ping
CP
Lu
D003001:Cloning, Molecular; D006867:Hydrolases / Q000235:genetics* / Q000378:metabolism; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D017011:Streptococcus suis / Q000201:enzymology* / Q000235:genetics
D011994:Recombinant Proteins; D006867:Hydrolases; C019723:arginine deiminase