Characterization and pathogenicity of the zinc metalloprotease empA of Vibrio anguillarum expressed in Escherichia coli.
The extracellular zinc metalloprotease (EmpA) is a putative pathogenic factor involved in the invasive process of the fish pathogen Vibrio anguillarum. It is synthesized as a 611-amino-acid preproprotease. The gene encoding EmpA (empA) has already been cloned and sequenced. In this study, empA was inserted into pET24d(+) and expressed in Escherichia coli BL21(DE3). Recombinant EmpA with His-tag was purified in a single step with a His-binding Ni-affinity column to a purity >95%. In addition, proteolytic activity, cytotoxicity, fish pathogenicity, and solubility of the recombinant protein were determined.
Yang
Hui
H
Department of Marine Biology, College of Marine Life Sciences, Ocean University of China, 5 Yushan Road, Qingdao 266003, PRC.
Chen
Jixiang
J
Yang
Guanpin
G
Zhang
Xiao-Hua
XH
Li
Yun
Y
Wang
Min
M
D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002460:Cell Line; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005393:Fish Diseases / Q000382:microbiology; D005432:Flounder / Q000382:microbiology; D015870:Gene Expression; D007928:Lethal Dose 50; D045726:Metalloproteases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016019:Survival Analysis; D014733:Vibrio / Q000201:enzymology* / Q000378:metabolism / Q000472:pathogenicity*; D014735:Vibrio Infections / Q000382:microbiology / Q000662:veterinary; D037521:Virulence Factors / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
D001426:Bacterial Proteins; D011994:Recombinant Proteins; D037521:Virulence Factors; C500552:EmpA protein, Vibrio anguillarum; D045726:Metalloproteases