Identification of genes induced in vivo during Klebsiella pneumoniae CG43 infection
Identification of genes induced in vivo during Klebsiella pneumoniae CG43 infection
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DOI：
10.1128/iai.69.11.7140-7145.2001
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发表时间：
2001-11-01
影响因子：
3.1
通讯作者：
Chang, HY
中科院分区：
文献类型：
--
作者：
Lai, YC;Peng, HL;Chang, HY
采用一种新的体内表达技术（IVET）鉴定肺炎克雷伯菌CG 43基因在感染BALB/c小鼠过程中特异性表达。IVET采用了一种UDP葡萄糖焦磷酸化酶（galU）缺陷型K. pneumoniae，其不能利用半乳糖和合成荚膜多糖，如通过其对BALB/c小鼠的低毒力和在麦氏半乳糖琼脂上的白色非粘液样菌落形态所证明的。通过使用功能性galU基因作为报告基因，RT 1启动子可以使galU突变体具有毒性，同时保持白色非粘液样集落表型。通过体内筛选共获得20个不同的序列。其中5个基因在其他病原体中已被鉴定为毒力相关基因，而另外5个具有特征性功能的基因参与营养吸收的调节和运输、类异戊二烯的生物合成和蛋白质折叠。其他10个序列没有已知的功能。我们还证明，20个IVE基因中的2个在缺铁的情况下启动，而另外5个基因的表达在超氧化物发生剂百草枯的存在下被激活。
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 RT, 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.