Identification of Mycobacterium tuberculosis RNAs synthesized in response to phagocytosis by human macrophages by selective capture of transcribed sequences (SCOTS)
Identification of Mycobacterium tuberculosis RNAs synthesized in response to phagocytosis by human macrophages by selective capture of transcribed sequences (SCOTS)
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DOI：
10.1073/pnas.96.20.11554
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发表时间：
1999-09-28
影响因子：
11.1
通讯作者：
Clark-Curtiss, JE
中科院分区：
文献类型：
--
作者：
Graham, JE;Clark-Curtiss, JE
建立了一种广泛适用的阳性cDNA筛选方法，用于鉴定结核分枝杆菌在培养的人原代巨噬细胞吞噬作用下合成的RNA，（替代sigma因子），aceA（异柠檬酸裂解酶），ponA（I类青霉素结合蛋白），PKS 2（聚酮合酶），uvrA（UvrABC内切核酸酶）和ctpV（推定的阳离子转运蛋白）的cDNA从巨噬细胞生长的细菌获得，ORF Rv 3070、Rv 3483 c、Rv 0903 c的cDNA（编码推定的细菌双组分转录激活因子），和mce 1毒力操纵子的Rv 0170也从吞噬的杆菌中获得。这些基因组区域的cDNA不能从在实验室肉汤中生长的大约1,000倍的细菌中获得。本文所述的方法已鉴定出M.结核病基因在宿主相互作用中的表达，将允许研究各种微生物中的基因表达，包括在自然疾病状态下与人类组织相互作用产生的表达。
A widely applicable, positive cDNA selection method was developed to identify RNAs synthesized by Mycobacterium tuberculosis in response to phagocytosis by cultured human primary macrophages, cDNAs for sigE and sigH (alternative sigma factors), aceA (isocitrate lyase), ponA (class I penicillin-binding protein), pks2 (polyketide synthase), uvrA (UvrABC endonuclease), and ctpV (putative cation transporter) were obtained from macrophage-grown bacteria, cDNAs for ORFs Rv3070, Rv3483c, Rv0903c (encoding a putative bacterial two-component transcriptional activator), and Rv0170 of the mce1 virulence operon also were obtained from phagocytized bacilli. cDNAs for these genomic regions were not obtained from approximately 1,000-fold more bacteria grown in laboratory broth. Methods described here, which have identified M. tuberculosis genes expressed in response to host interaction, will allow the study of gene expression in a variety of microorganisms, including expression resulting from interaction with human tissues in natural disease states.