Grange Immunol. Phenotypically, they exhibited attenuated but distinct virulence in mice. Error bars representing ±SD are shown in (D) and are smaller than symbols in (B). This revealed that the nuoG mutation did not induce a major change in the proteome (Figure S8). GFP-expressing BCG and M. smegmatis were generated by subcloning the enhanced GFP gene (Clontech, http://www.clontech.com/) into the mycobacterial episomal expression vector pMV261. The 247 bp remnant and its probable original counterpart in H37Rv are shaded. Conversely, deletion of nuoG in M. tuberculosis ablated its ability to inhibit macrophage apoptosis and significantly reduced its virulence in mice. The RT-PCR data of the transcription levels of selected genes with promoter mutations were subjected to statistical analysis. MRA_1772 has a 1 bp insertion which induced a premature termination relative to its H37Rv ortholog Rv1759c encoding a fibronectin binding PE-PGRS family protein [49], causing it to be 2398 bp shorter than the original stop codon. Arachne [63] and Phrap (www.phrap.org) were used for sequence assembly. The genotype of Mycobacterium tuberculosis is an important factor in the interaction between the bacilli and the host, as well as in the preclinical study of drugs and vaccines. After turning cultured positive, 0.5 mL of each culture was inoculated into MGIT 7 mL tubes with SIRE supplement and isoniazid, rifampicin or streptomycin, then the tubes were scanned into the system to be cultured and the respective drug sensitivity was determined and reported. D.A. <<030C923D67FA0F4AADD28A65A04E6A6D>]>> Howard Hughes Medical Institute, Albert Einstein College of Medicine, Bronx, New York, United States of America The H37Ra was grown in Middlebrook 7H9 liquid medium with ADC (albumin-dextrose-catalase) supplement to late log phase for preparation of high-molecular weight genomic DNA as described [62]. We determined the complete genomic sequence of H37Ra ATCC25177 and compared that with its virulent counterpart H37Rv and a clinical isolate CDC1551. Extraction of Mycobacterium tuberculosis DNA: a question of containment. The MRA_2376 is identical to MRA_2373, and the same gene duplication is observed in CDC1551 (MT2419 and MT2422). ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. The 6 real “H37Rv-specific” variations (i.e., those that are different from both H37Ra and CDC1551) are likely acquired during long-term repeated in vitro cultivation of H37Rv over the years but unrelated to the virulence property. Highly confident SNPs which had been confirmed in previous research were used to illustrate the separate resistance (Tables 1, 2). (2015). The Mtb nuoG mutant induced significantly more apoptosis than the complemented strain or the wild-type Mtb in human and mouse macrophages (Figure 3B and 3C). KV, WRJ, SAP, and VB analyzed the data. (2011). Apart from lpdA, other genes with promoter region mutations may also be involved in virulence attenuation since those genes in H37Ra had different expression patterns compared to H37Rv (Figure 2). Yes Role of M. tuberculosis RD-1 region encoded secretory proteins in protective response and virulence. Shanghai-MOST Key Laboratory of Health and Disease Genomics, Chinese National Human Genome Center at Shanghai, Shanghai, China, Affiliation Lungs and spleens were collected and fixed in 4% formaldehyde for 72 h, followed by paraffin embedded sectioning and H&E stain.

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