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Infection and Immunity, November 2001, p. 7140-7145, Vol. 69, No. 11
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.11.7140-7145.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Identification of Genes Induced In Vivo during Klebsiella pneumoniae CG43 Infection

Yi-Chyi Lai,1 Hwei-Ling Peng,2 and Hwan-You Chang1,*

Department of Life Science and Institute of Biotechnology, National Tsing Hua University,1 and Department of Biological Science and Technology, National Chiao Tung University,2 Hsin Chu, Taiwan, Republic of China

Received 16 January 2001/Returned for modification 20 March 2001/Accepted 20 July 2001

A novel in vivo expression technology (IVET) was performed to identify Klebsiella pneumoniae CG43 genes that are specifically expressed during infection of BALB/c mice. The IVET employed a UDP glucose pyrophosphorylase (galU)-deficient mutant of K. pneumoniae which is incapable of utilizing galactose and synthesizing capsular polysaccharide, as demonstrated by its low virulence to BALB/c mice and a white nonmucoid colony morphology on MacConkey-galactose agar. By using a functional galU gene as the reporter, an IVE promoter could render the galU mutant virulent while maintaining the white nonmucoid colony phenotype. A total of 20 distinct sequences were obtained through the in vivo selection. Five of them have been identified previously as virulence-associated genes in other pathogens, while another five with characterized functions are involved in regulation and transportation of nutrient uptake, biosynthesis of isoprenoids, and protein folding. No known functions have been attributed to the other 10 sequences. We have also demonstrated that 2 of the 20 IVE genes turn on under iron deprivation, whereas the expression of another five genes was found to be activated in the presence of paraquat, a superoxide generator.

* Corresponding author. Mailing address: Department of Life Science, National Tsing Hua University, 101 Kuan-Fu Road, 2nd Sec., Hsin Chu, Taiwan, Republic of China. Phone: 886-3-5742910. E-mail: hychang{at}life.nthu.edu.tw.

Infection and Immunity, November 2001, p. 7140-7145, Vol. 69, No. 11
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.11.7140-7145.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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