行政院國家科學委員會專題研究計畫期中進度報告:以標籤式突變法分析Salmonella choleraesuis活體內毒力因子(1/3)
Date Issued
2003
Date
2003
Author(s)
張照夫
DOI
912313B002386
Abstract
Salmonella choleraesuis is a facultative intracellular pathogen that causes a
septicemic disease of pigs. S. choleraesuis is host-adapted for swine. S. choleraesuis
occasionally also causes human infection, so it is a zoonotic pathogen. Although S.
choleraesuis causes an economic loss in swine industry, the information about the
virulence genes of this organism is very limited. The hypothesis is that there are many
proteins which are only present when the bacteria are growing in the animal host (in
vivo) and not when the pathogen is grown under laboratory conditions (in vitro), and
that these proteins play a critical role in the disease process. The goal is to identify
genes encoding potential virulence factors that are expressed only in vivo in the swine
host. These proteins may prove to be important in the pathogenesis of S. choleraesuis
infection in pigs. Generation of the S. choleraesuis mutant bank was the purpose of
the first year project. A series of identical membranes for dot blot hybridizations was
prepared by transferring of 282 plasmids onto Hybond N+ membranes using the
Bio-dot Microfiltration Apparatus. To test whether the hybridization signals resulted
from cross-hybridization between tags, amplified tags from 94 plasmids were used to
probe a dot blot of 282 plasmids, which included the 94 used to generate the probe.
There were cross-hybridization to the probe. These plasmids were deleted. The same
method was repeatedly for the other plasmid. Cross-hybridization was continued but
the amount of plasmid was used to probe were 48, 24 and 8. Eighty-two pUT
mini-Tn5Km2 plasmids containing unique tag were found. A set of 48 plasmids
containing strongly-hybridizing tags was repeatedly used to construct 20 libraries of S.
choleraesuis.
Subjects
Salmonella choleraesuis
signature-tagged mutagenesis
in vivo virulence
factor
factor
SDGs
Publisher
臺北市：國立臺灣大學獸醫學系暨研究所
Type
report
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