Browsing by Author "Fung, Chang-Phone"

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Characterization of the type 3 fimbriae with different MrkD adhesins: Possible role of the MrkD containing an RGD motif
(2006-11-24)
Huang, Ying-Jung
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Wu, Chien-Chen
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Chen, Mei-Chen
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Fung, Chang-Phone
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Peng, Hwei-Ling
Four novel mrkD alleles namely mrkD(V1), mrkD(V2), mrkD(V3), and mrkD(V4) were identified in seventeen Klebsiella pneuntoniae meningitis strains using PCR-RFLP and sequence determination. Comparative analysis revealed a most variable region containing an RGD motif in the receptor domain of MrkD(V3). In order to determine if the sequence confers the K pneumoniae mrkD(V3) the highest level of the fimbrial activity, a type 3 fimbriae display system was constructed in Escherichia coli. The E coli JM 109[pmrkABCD(V3)F] displaying meshwork-like fimbriae also had the most fimbrial activity, supporting a possible role of the varied sequences. In a dose-dependent manner, the GRGDSP hexapeptide appeared to inhibit the adhesion of the E. coli JM109[pmrkABCD(V3)F] to HCT-8, an ileocecal epithelial cell line. In addition, the adhesion activity was reduced by the addition of anti-alpha 5 beta 1 integrin monoclonal antibody, indicating that the RGD containing region in MrkD(V3) is responsible for the binding of type 3 fimbriae to integrin. (c) 2006 Elsevier Inc. All rights reserved.
Regulation of the Klebsiella pneumoniae Kpc fimbriae by the site-specific recombinase Kpcl
(2010-07-01)
Wu, Chien-Chen
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Huang, Ying-Jung
;
Fung, Chang-Phone
;
Peng, Hwei-Ling
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生物科技學系
In the genome of Klebsiella pneumoniae NTUH-K2044, nine fimbrial gene clusters were identified. Besides type 1 and type 3 fimbriae, the others are novel and were named Kpa, Kpb, Kpc, Kpd, Kpe, Kpf and Kpg fimbriae. Prevalence analysis among 105 K. pneumoniae clinical isolates revealed that the kpc genes were highly associated with the K1 serotype isolates. Induced expression of the recombinant kpcABCD genes in Escherichia colt resulted in Kpc fimbriation and increased biofilm formation. A putative site-specific recombinase encoding gene kpcl and a 302 bp intergenic DNA flanked by 11 bp inverted repeats, namely kpcS, were identified in the upstream region of the kpcABCD genes. Using LacZ as the reporter, a dramatic difference in promoter activity of kpcS in two different orientations was observed and accordingly assigned as ON and OFF phase. kpcl expression was found to be able to invert kpcS in trans from phase ON to OFF and vice versa. Using the two-plasmid system, expression of kpcA, encoding the major component of the Kpc fimbriae, could be observed upon the induced expression of kpcl. These results indicate that Kpcl is involved in the regulation of Kpc fimbriation in a phase-variable manner.