| Summary: | 碩士 === 中國醫藥大學 === 醫學研究所碩士班 === 95 === Klebsiella pneumoniae (K. pneumoniae) infection results in various illnesses including liver abscess in Taiwan. The capsule of K. pneumoniae is generally considered as a virulence factor. However, in our previous study, hypomucoid K. pneumoniae was highly invasive in diabetic mice and caused a high morbidity and mortality. To study if any virulence genes other than mucoviscosity are present in hypomucoid K. pneumoniae strains, 3 spots were revealed from the comparison of hypomucoid KP 1084 versus hypermucoid KP 1112 with by two-dimensional gel electrophoresis. Employing bioinformatics method, two protein spots were identified as chaperones DnaK and GroEL, which were shown to be able to bind with carbohydrate lyase “GatY”. To find the genes that may be crucial for the pathogenesis of K. pneumoniae-induced liver abscess, subtraction hybridization was employed. The substracted DNA fragments could discriminate different clusters of K. pneumoniae. By using BLASTx, the potential proteins could be identified. These included glycosyltransferase, carbohydrate lyase “GatY” and transposase that stand for K. pneumoniae cluster AA’, integrase that stands for clusterA’, and certain protein domains, such as secretory lipase and dienelactone hydrolase that may stand for cluster CDD’.
Furthermoe, we used bioinformatics method to align and compare, between K. pneumoniae and E. coli, the differences of genes for carbohydrate utilization, stress responses and osmolar regulation. The result showed that the sequence of loop1 on OmpA of K. pneumoniae was 15 bases longer than that of E coli, and most of the proteins translated from the longer sequence s were valine, leucine, glutamate. The sequence of loop3 on OmpA of K. pneumoniae was 12 bases longer than that of E coli, and most from the longer sequence were alanine and glycine. These differences may affect the binding ability of OmpA. This study may lead to the elucidation of the pathogenesis of K. pneumoniae.
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