[MrkD adhesin of Klebsiella pneumoniae expression, purification and analysis of adhesive activity].
OBJECTIVE
The mrkD gene encodes the adhesin which mediates Klebsiella pneumoniae to adhere human respiratory tissue. We aimed to analyze the adhesion mechanism and adhesion block function of MrkD adhesin.


METHODS
The recombinant glutathione-S-transferase (GST)-tagged adhesive protein (MrkD) was expressed in E. coli and was purified to homogeneity using GST affinity chromatography. The GST tag was cut by thrombin to obtain the MrkD protein that was identified by SDS-PAGE and Western blot. The adhesive activity of MrkD was examined by adhesive experiments and the binding site was observed by laser confocal microscopy.


RESULTS
The adherent activity of Klebsiella pneumoniae was significantly inhibited by the MrkD. These experimental data demonstrated that the MrkD inhibited the adhesion of Klebsiella pneumoniae.


CONCLUSIONS
Our results suggest that MrkD adhesin contains the adhesion epitopes. The future work will be carried out to identify the epitopes and characterize them, then to optimize the combination presentation of these epitopes to develop an efficient vaccine for Klebsiella pneumoniae.
Li
Yang
Y
College of Animal Science and Veterinary Medicine, Jilin University, Changchun 130062, China. myth0317@126.com
Han
Wenyu
W
Lei
Liancheng
L
Li
Zhijie
Z
Shi
Lei
L
D018829:Adhesins, Bacterial / Q000032:analysis / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism* / Q000502:physiology; D000268:Adhesiveness; D001422:Bacterial Adhesion / Q000502:physiology*; D001426:Bacterial Proteins; D019411:Clinical Laboratory Techniques; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D036781:Fimbriae Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism* / Q000502:physiology; D015870:Gene Expression; D007711:Klebsiella pneumoniae / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; C058768:Klebsiella pneumoniae type 3 fimbrial adhesin; D011994:Recombinant Proteins; D036781:Fimbriae Proteins