*Target (protein/gene name): 3-oxoacyl-[acyl-carrier-protein] reductase / fabG1
*NCBI Gene # or RefSeq#: 886551
*Protein ID (NP or XP #) or Wolbachia#: NP_215999.1
*Organism (including strain): Mycobacterium tuberculosis H37Rv
Etiologic Risk Group (see link below): Risk Group 3
*Background/Disease Information (sort of like the Intro to your Mini Research Write up):
Tuberculosis is a potentially fatal disease that is caused by the bacteria Mycobacterium tuberculosis. In the past, tuberculosis was the leading cause of death in the United States. Although this is no longer the case due to vaccinations and immunizations, the threat of tuberculosis is still a reality. African-Americans, pregnant women, and internal travelers are all at higher risks to contracting tuberculosis based on statistical data. Complications with eradicating tuberculosis include the development of drug-resistance from poor quality drugs or people not completing treatment. Tuberculosis can also stay inactive in the body and become activated causing the normal symptoms of tuberculosis to arise. This has been termed latent tuberculosis infection. [1]
Essentiality of this protein:
-Gene/Ortholog: mtu1373 (OG4_10098); Phenotype: essential; Source study: nmpdr

-Gene/Ortholog: eco1057 (OG4_10098); Phenotype: essential; Source study: gerdes

Complex of proteins?: No
Druggable Target: Yes, 0.7 druggability index

*EC#: 1.1.1.100
Link to BRENDA EC# page: http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.1.1.100 [2]
-- Show screenshot of BRENDA enzyme mechanism schematic
reaction schematic.png

Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): Spectrophotometric; Reagents: 100 mM Potassium phosphate (pH 7.0), NADPH (100 uM), acetoacetyle-N-acetylcysteamine (5mM)
-- link to Sigma (or other company) page for assay or assay reagents (substrates)
Potassium phosphate, NADPH, acetoacetyle-N-acetylcysteamine [3]
-- link (or citation) to paper that contains assay information
For same protein in different organism (Persea americana)
Sheldon, P.S.; Kekwick, R.G.O.; Sidebottom, C.; Smith, C.G.; Slabas, A.R., 3-Oxoacyl-(acyl-carrier protein) redictase from avocado (Persia americana) fruit mesocarp. Biochem. J. 1990, 271, 713-720. [4]
-- List cost and quantity of substrate reagents and supplier
Potassium phosphate - $20.80
NADPH - $99.90
acetoacetyle-N-acetylcysteamine - $70.10

Structure Available (PDB or Homology model)
-- PDB # or closest PDB entry if using homology model: 1UZN [5]
-- For Homology Model option: N/A
---- Show pairwise alignment of your BLASTP search in NCBI against the PDB: N/A
---- Query Coverage: N/A
---- Max % Identities: N/A
---- % Positives: N/A
---- Chain used for homology: N/A

Current Inhibitors: 14 in the species Plasmodium falciparum but none in Mycobacterium tuberculosis on The Binding Database
Expression Information (has it been expressed in bacterial cells): Yes, in E. coli [6]
Purification Method: Column Chromatography - "Induced recombinant bacteria were washed with cold 50 mM potassium phosphate buffer pH 7±8 (buffer A), and resuspended in 4 ml of the same buffer supplemented with 500 mM NaCl and 5 mMimidazole. Cells were broken by one freezing}thawing cycle at®70 °C, in the presence of protease inhibitors [aprotinin, soybean trypsininhibitor, TLCK (N-p-tosyl-lysine chloromethyl ketone), pepstatin, leupeptin and PMSF] and 0±5 mg lysozyme ml−". Nucleic acids were removed by DNase I (5 lg ml−") and RNase A (10 lg ml−") treatments in the presence of MgCl# (10 mM) at 4 °C for 15 min. After centrifugation at 44000 g for 15 min, the supernatant supplemented with 10% (v}v) glycerol was applied to a Ni-NTA Agarose (Qiagen) column (0±5 ml bed volume). After extensive washes by 5 mM then 50 mM imidazole in buffer A supplemented with 500 mM NaCl, MabA was eluted with 175 mM imidazole in buffer A supplemented with 500 mM NaCl. Fractions containing the His-tagged MabA (H-MabA) were identi®ed by SDS-PAGE, pooled, dialysed twice against 50 vols 50% (v}v) glycerol in 50 mM potassium phosphate buffer, pH 7±2, and stored at ®20 °C." (Same paper as expression linked above). [6]
Image of protein (PyMol with features delineated and shown separately):
1UZN.png
*Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):
MTATATEGAKPPFVSRSVLVTGGNRGIGLAIAQRLAADGHKVAVTHRGSGAPKGLFGVEVDVTDSDAVDRAFTAVEEHQG
PVEVLVSNAGLSADAFLMRMTEEKFEKVINANLTGAFRVAQRASRSMQRNKFGRMIFIGSVSGLWGIGNQANYAASKAGV
IGMARSIARELSKANVTANVVAPGYIDTDMTRALDERIQQGALQFIPAKRVGTPAEVAGVVSFLASEDASYISGAVIPVD
GGMGMGH

*length of your protein in Amino Acids: 247 residues
Molecular Weight of your protein in kiloDaltons using the Expasy ProtParam website: 25.7192 kDa [7]
Molar Extinction coefficient of your protein at 280 nm wavelength: 9970
TMpred graph Image (http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.
Graph.gif[8]
*CDS Gene Sequence (paste as text only):
GTGACTGCCACAGCCACTGAAGGGGCCAAACCCCCATTCGTATCCCGTTCAGTCCTGGTTACCGGAGGAAA
CCGGGGGATCGGGCTGGCGATCGCACAGCGGCTGGCTGCCGACGGCCACAAGGTGGCCGTCACCCACCG
TGGATCCGGAGCGCCAAAGGGGCTGTTTGGCGTCGAATGTGACGTCACCGACAGCGACGCCGTCGATCGC
GCCTTCACGGCGGTAGAAGAGCACCAGGGTCCGGTCGAGGTGCTGGTGTCCAACGCCGGCCTATCCGCGG
ACGCATTCCTCATGCGGATGACCGAGGAAAAGTTCGAGAAGGTCATCAACGCCAACCTCACCGGGGCGTTC
CGGGTGGCTCAACGGGCATCGCGCAGCATGCAGCGCAACAAATTCGGTCGAATGATATTCATAGGTTCGGT
CTCCGGCAGCTGGGGCATCGGCAACCAGGCCAACTACGCAGCCTCCAAGGCCGGAGTGATTGGCATGGCC
CGCTCGATCGCCCGCGAGCTGTCGAAGGCAAACGTGACCGCGAATGTGGTGGCCCCGGGCTACATCGACA
CCGATATGACCCGCGCGCTGGATGAGCGGATTCAGCAGGGGGCGCTGCAATTTATCCCAGCGAAGCGGGT
CGGCACCCCCGCCGAGGTCGCCGGGGTGGTCAGCTTCCTGGCTTCCGAGGATGCGAGCTATATCTCCGGTG
CGGTCATCCCGGTCGACGGCGGCATGGGTATGGGCCACTGA
[9]
*GC% Content for gene: 64.4% [10]
*CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only):
*GC% Content for gene (codon optimized):

Do Not Need this info for Spring (but still copy these lines to your Target page for now)
Primer design results for pNIC-Bsa4 cloning (list seqeunces of all of your ~40 nt long primers):
(link to DNA Works output text file - that should be saved in your Google Docs folder after you did the primer design protocol)
-- Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to.

Primer design results for 'tail' primers (this is just 2 sequences):

References:
1.http://www.cdc.gov/tb/publications/factsheets/general/tb.htm
2. http://www.brenda-enzymes.org/php/result_flat.php4?ecno=1.1.1.100
3. http://www.sigmaaldrich.com/sigma-aldrich/home.html
4. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1149621/pdf/biochemj00172-0147.pdf
5. http://www.pdb.org/pdb/explore/explore.do?structureId=1UZN
6. http://mic.sgmjournals.org.ezproxy.lib.utexas.edu/content/148/4/951.full.pdf+html
7. http://web.expasy.org/protparam/
8. http://www.ch.embnet.org/software/TMPRED_form.html
9. http://www.ncbi.nlm.nih.gov/gene/886551
10. http://www.sciencebuddies.org/science-fair-projects/project_ideas/Genom_GC_Calculator.shtml
11. http://tdrtargets.org/