TABLE 1.
The bTB ORFs selected for recombinant expression tests.
| Expression host | Soluble expression | Purification yield | ||
|---|---|---|---|---|
| Cell wall and cell processes | ||||
| Mb0923 | Outer membrane protein Omp A | E. coli & Insect | +++ | +++ |
| Mb0485 | Iron-regulated heparin binding hemagglutinin hbha | E. coli & Insect | +++ | +++ |
| Mb0419c | Probable glutamine-binding lipoprotein glnh (glnbp) | E. coli & Insect | ++ | + |
| Mb3840 | Exported repetitive protein precursor PirG | E. coli & Insect | +++ | + |
| Mb0348 | Isoniazid inductible gene protein iniB | E. coli & Insect | +++ | +++ |
| Mb2293 | Probable lipoprotein lppN | E. coli | +++ | +++ |
| Mb2807c | Probable lipoprotein lppU | Insect | +++ | +++ |
| Mb0598c | Probable lipoprotein lpqN | E. coli & Insect | +++ | +++ |
| Mb1260 | Probable lipoprotein lpqX | Insect | +++ | +++ |
| Mb1568c | Probable lipoprotein lprI | E. coli & Insect | +++ | +++ |
| Mb1891c | Alanine and proline rich secreted protein APA | E. coli & Insect | +++ | +++ |
| Mb3905 | 6 kda early secretory antigenic target esxa (Esat-6) | E. coli & Insect | +++ | +++ |
| Mb3904 | 10 kda culture filtrate antigen esxb (lhp) (cfp10) | E. coli & Insect | +++ | +++ |
| Mb0296 | Low molecular weight antigen 7 esxh (10 kda antigen) (cfp-7) | Insect | +++ | +++ |
| Mb0295 | Esat-6 like protein esxg | Insect | +++ | +++ |
| Mb1229 | Esat-6 like protein esxk (Esat-6 like protein 3) | E. coli & Insect | +++ | +++ |
| Mb1230 | Putative Esat-6 like protein esxi (Esat-6 like protein 4) | Insect | +++ | +++ |
| Mb1820 | Esat-6 like protein esxm | E. coli & Insect | +++ | +++ |
| Mb1821 | Putative Esat-6 like protein esxn (Esat-6 like protein 5) | Insect | +++ | ++ |
| Mb3475c | Esat-6 like protein esxu | E. coli & Insect | +++ | +++ |
| Mb3911c | Proteolytic substrate protein espb | E. coli & Insect | ++ | ++ |
| Mb0674 | Possible ribonucleotide-transport ATP-binding protein | E. coli & Insect | +++ | ++ |
| Mb1036 | Probable resuscitation-promoting factor rpfb | E. coli & Insect | ++ | - |
| Mb1445 | Aminoglycosides/tetracycline-transport integral membrane protein | E. coli & Insect | ++ | - |
| Mb3070 | Probable FeIII-dicitrate-binding periplasmic lipoprotein fecb | E. coli | ++ | ++ |
| Mb3338 | Acid phosphatase sapm | E. coli & Insect | ++ | ++ |
| Mb2653c | Probable conserved transmembrane protein | E. coli & Insect | - | - |
| Mb3646c | Esx-1 secretion-associated protein, espa | E. coli & Insect | +++ | + |
| Mb3645c | Esx-1 secretion-associated protein espc | E. coli & Insect | +++ | +++ |
| Mb3644c | ESX-1 secretion-associated protein espd | E. coli & Insect | - | - |
| Mb2002c | Immunogenic protein mpt64 (antigen mpt64/mpb64) | E. coli & Insect | +++ | +++ |
| Mb1762 | Probable conserved transmembrane protein | E. coli & Insect | 0 | ++ |
| Information pathway related proteins | ||||
| Mb0055 | Single-strand binding protein Ssb | E. coli & Insect | + | + |
| Mb0671 | 50s ribosomal protein l7/l12 rplL (sa1) | E. coli & Insect | +++ | +++ |
| Mb0704 | Probable iron-regulated elongation factor tu tuf | E. coli & Insect | ++ | + |
| Mb0670 | 50s ribosomal protein l10 rplJ | E. coli & Insect | +++ | ++ |
| Mb1656 | 30s ribosomal protein s1 rpsA | E. coli & Insect | +++ | +++ |
| Virulence related proteins | ||||
| Mb3441 | Possible antitoxin VapB47 | Insect | +++ | +++ |
| Mb2891 | Toxin RelG | E. coli & Insect | +++ | - |
| PE/PPE family proteins | ||||
| Mb2548 | PE family PE26 | Insect | + | + |
| Rv3622c | PE family protein PE32 | E. coli & Insect | +++ | +++ |
| Mb0293 | PE family protein PE5 | E.coli & Insect | +++ | +++ |
| Mb0294 | PE family protein PE4 | E.coli & Insect | +++ | +++ |
| Mb3902 | PE family-related protein PE35 | E.coli & Insect | +++ | +++ |
| Mb3922c | PE family protein PE36 | E.coli | +++ | +++ |
| Mb3504 | PE family protein PE31 | Insect | +++ | +++ |
| Mb0940c | PE family protein PE7 | Insect | +++ | +++ |
| Mb1835 | PE family protein PE20 | Insect | +++ | +++ |
| Mb1421 | PE family protein PE15 | E.coli & Insect | +++ | +++ |
| Mb3505 | PE family protein PPE 60 | E.coli & Insect | +++ | +++ |
| Mb1069c | PE family protein PE 8 | E.coli & Insect | +++ | +++ |
| Mb2457c | PE family protein PE 25 | E.coli & Insect | +++ | +++ |
| Mb1202c | PE family protein | E.coli & Insect | +++ | +++ |
| Mb3903 | PPE family protein PPE68 | E.coli & Insect | +++ | +++ |
| Mb3921c | PPE family-related protein PPE69 | E.coli & Insect | +++ | +++ |
| Mb0939c | PPE family protein PPE 14 | E.coli & Insect | +++ | +++ |
| Mb1836 | PPE family protein PPE 31 | E.coli & Insect | +++ | +++ |
| Mb1068c | PPE family protein PPE15 | E.coli & Insect | +++ | +++ |
| Mb1422 | PPE family protein PPE 20 | Insect | +++ | +++ |
| Mb2456c | PPE family protein PE41 | E.coli & Insect | +++ | +++ |
| Mb1228 | PPE family protein PPE 18 | E.coli & Insect | +++ | +++ |
| Mb1200c | PPE17 (part) | E.coli & Insect | +++ | +++ |
| Mb1201c | PPE 17 (part) | E.coli & Insect | +++ | +++ |
| Intermediary metabolism & respiration related proteins | ||||
| Mb3871 | Bacterioferritin BfrB | E.coli | +++ | +++ |
| Mb2981 | Possible glycosyl transferase | E.coli & Insect | ++ | + |
| Mb0130 | probable serine protease pepA | E.coli & Insect | ++ | ++ |
| Mb1272 | Probable malate dehydrogenase mdh | E.coli & Insect | ++ | - |
| Mb0977 | Probable succinyl-CoA synthetase (a chain) sucD | E.coli & Insect | ++ | ++ |
| Mb3652 | Inorganic pyrophosphatase PPA | E.coli & Insect | +++ | |
| Mb1129c | Fructose 1,6-bisphosphatase glpX | Insect | ++ | - |
| Mb3412 | Diterpene synthase | E.coli & Insect | ++ | ++ |
| Conserved hypotheticals & unknown proteins | ||||
| Mb2315c | Hypothetical protein | Insect | + | - |
| Mb3935c | Putative Esat-6 like protein esxf (Esat-6 like protein 13) | E. coli & Insect | +++ | +++ |
| Mb3920c | Possible Esat-6 like protein esxd | E. coli & Insect | +++ | +++ |
| Mb3934c | Putative Esat-6 like protein esxe (Esat-6 like protein 12) | E. coli & Insect | +++ | +++ |
| Mb1066c | Putative Esat-6 like protein esxI (Esat-6 like protein 1) | Insect | +++ | +++ |
| Mb1067c | Esat-6 like protein esxj | E. coli & Insect | +++ | +++ |
| Mb2375c | Putative Esat-6 like protein esx0 (Esat-6 like protein 6) | Insect | + | + |
| Mb3042c | Esat-6 like protein esxq (tb12.9) (Esat-6 like protein 8) | Insect | + | + |
| Mb3045c | Secreted Esat-6 like protein esxr (Esat-6 like protein 9) | E.coli & Insect | +++ | +++ |
| Mb3046c | Esat-6 like protein esxs | E.coli & Insect | +++ | +++ |
| Mb3474c | Putative Esat-6 like protein esxt | E.coli & Insect | +++ | +++ |
| Mb3919c | Esat-6 like protein esxc (Esat-6 like protein 11) | Insect | + | - |
| Mb0959 | Periplasmic phosphate-binding lipoprotein PstS1 | E. coli & Insect | +++ | +++ |
| Mb1858 | Conserved protein with fha domain, gara | E. coli & Insect | ++ | ++ |
| Mb1868c | Malate synthase G GlcB | E. coli & Insect | ++ | ++ |
| Mb1943c | Catalase-peroxidase-peroxynitritase T KatG | E. coli & Insect | ++ | ++ |
| Mb2006c | Probable cutinase precursor CFP21 | E. coli & Insect | ++ | +++ |
| Mb2057c | Stress protein induced by anoxia | E. coli & Insect | ++ | +++ |
| Mb2244 | Glutamine synthetase GlnA1 | E. coli & Insect | ++ | +++ |
| Mb2898 | Cell surface lipoprotein Mpt83 (lipoprotein P23) | E. coli & Insect | ++ | +++ |
| Mb2900 | Major secreted immunogenic protein Mpt70 | E. coli & Insect | ++ | ++ |
| Mb0169 | Conserved protein TB18.5 | E. coli & Insect | + | + |
| Mb0418c | Serine/threonine-protein kinase PknG | E. coli & Insect | +++ | +++ |
| Mb2477c | Probable resuscitation-promoting factor RpfE | E. coli & Insect | ++ | - |
| Mb1319 | Conserved protein | E. coli & Insect | ++ | ++ |
| Mb1596 | Involved in biotin biosynthesis | E. coli & Insect | ++ | ++ |
| Mb2656 | Universal stress protein family protein TB31.7 | E. coli & Insect | ++ | ++ |
| Mb3046c | Esat-6 like protein EsxS | E. coli & Insect | ++ | ++ |
| Mb2982c | Possible glycosyl transferase | E. coli & Insect | ++ | + |
| Mb0455c | Cyclopropane fatty acid synthase | E. coli & Insect | +++ | ++ |
| Mb2054c | pfkb | E. coli & Insect | +++ | +++ |
| Mb3157c | devR | E. coli & Insect | +++ | +++ |
| Mb2970c | Probable conserved lipoprotein LppX | E. coli & Insect | ++ | |
| Mb0891 | Possible resuscitation-promoting factor rpfA | E. coli & Insect | +++ | ++ |
| Mb1916 | Probable resuscitation-promoting factor rpfC | E. coli & Insect | ++ | ++ |
| Mb2410 | Probable resuscitation-promoting factor rpfD | E. coli & Insect | ++ | ++ |
| Mb3274c | Two component sensory transduction transcriptional regulatory protein mtrA | E. coli & Insect | +++ | +++ |
| Mb0463c | Conserved protein | E. coli & Insect | +++ | +++ |
| Mb3641 | Hypothetical arginine and proline rich protein | E. coli & Insect | ++ | - |
| Mb0062 | Hypothetical protein | E. coli & Insect | ++ | ++ |
| Mb1843 | Conserved protein | E. coli & Insect | ++ | ++ |
| Mb3743c | Conserved protein | E. coli & Insect | +++ | +++ |
| Mb1833c | Conserved protein | Insect | +++ | +++ |
| Mb0854c | Conserved protein | E. coli & Insect | +++ | +++ |
| Mb2058 | Conserved protein Acg | E. coli & Insect | ++ | - |
| Mb0337c | hypothetical protein | E. coli & Insect | ++ | ++ |
| Mb2660c | Conserved protein | E. coli & Insect | ++ | - |
| Mb 2659 | hypoxic response protein 1 hrp1 | E. coli | +++ | +++ |
| Mb3707 | Probable bifunctional membrane-associated penicillin-binding protein 1a/1b pona2 | E. coli & Insect | ++ | + |
| Mb0014c | Transmembrane Serine/therorine protein Kinase-B (pknb) | E. coli & Insect | ++ | +++ |
| Mb0979 | Probable conserved Transmembrane protein | E. coli & Insect | ++ | +++ |
| Mb0448 | GROEL protein-2 | E. coli & Insect | ++ | +++ |
Soluble expression levels: +++ - strongest band on SDS-PAGE, ++ - among the stronger bands, + - visible band, − no visible band. Purification yields: +++ ∼1 mg/L, ++ ∼0.1 mg/L, + <0.1 mg/L, - not purified. Greyed boxes required wash and elution buffers with 0.1% sodium sarkosyl.