Dihyrofolate+Reductase+(Mycobacterium+tuberculosis)


 * Target (protein/gene name):**

Dihyrdofolate Reductase Mycobacterium Tuberculosis 1DF7


 * NCBI Gene # or RefSeq#:**

GI: 581362


 * Protein ID (NP or XP #) or Wolbachia#:**

1DF7


 * Organism (including strain):**

Mycobacterium Tuberculosis 1DF7


 * Etiologic Risk Group (see link below):**

Risk Group 3 (RG3)


 * Background/Disease Information (sort of like the Intro to your Mini Research Write up):**

Tuberculosis is a lethal and infectious disease caused the mycobacteria strain Mycobacterium tuberculosis. Tuberculosis usually affects the lungs, but it can also spread to the rest of the body. It is spread through the air, especially when people cough or sneeze. If tuberculosis is left untreated it could kill more than 50% of those infected.

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 * Essentiality of this protein:**

It is essential. []


 * Complex of proteins?:**

no


 * Druggable Target:**

Yes []


 * *EC#: **


 * Link to BRENDA EC# page:**


 * --** Show screenshot of BRENDA enzyme mechanism schematic

Figure. 1. BRENDA Enzyme mechanism schematic for Dihydrofolate reductase.


 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents):**

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 * -- link to Sigma (or other company) page for assay or assay reagents (substrates)**

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 * -- link (or citation) to paper that contains assay information**

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 * -- List cost and quantity of substrate reagents and supplier**

50 mM Potassium Phosphate Buffer, pH 6.5 at 25°C $**21.70**

0.11 mM ß-Nicotinamide Adenine Dinucleotide Phosphate, Reduced Form Solution **(Price not found on sigma)**

2.3 mM Dihydrofolic Acid Solution (DHFA) **(Price not found on sigma)**

0.1% (w/v) Bovine Serum Albumin Solution (BSA) **$193.00**


 * Structure Available (PDB or Homology model)**

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 * Current Inhibitors:**

Zn2+ Li+ Mg2+


 * Expression Information (has it been expressed in bacterial cells):**

Yes it is expressed in other bacteria cells. One such example is E. Coli.


 * Purification Method:**

Nickel or His tag purification.


 * Image of protein (PyMol with features delineated and shown separately):**

Figure 1. Dihydrofolate Reductase protein shown in cartoon. The orange spirals are helices and the blue arrows are beta sheets.


 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):**

MVGLIWAQATSGVIGRGGDIPWRLPEDQAHFREITMGHTIVMGRRTWDSLPAKVRPLPGRRNVVLSRQADFMASGAEVVG SLEEALTSPETWVIGGGQVYALALPYATRCEVTEVDIGLPREAGDALAPVLDETWRGETGEWRFSRSGLRYRLYSYHRS


 * length of your protein in Amino Acids**

159


 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website:**

17640 kiloDaltons


 * Molar Extinction coefficient of your protein at 280 nm wavelength:**

code coefficient   40450 Abs 0.1% (=1 g/l)  2.293, assuming all pairs of Cys residues form cystines

Ext. coefficient   40450 Abs 0.1% (=1 g/l)  2.293, assuming all Cys residues are reduced

code


 * TMpred graph Image ** ( @http://www.ch.embnet.org/software/TMPRED_form.html ). Input your amino acid sequence to it.




 * CDS Gene Sequence (paste as text only):**

code ATGGTGGGGCTGATCTGGGCTCAAGCGACATCGGGTGTCATCGGCCGCGGCGGCGACATCCCCTGGCGCT TGCCCGAGGACCAGGCGCATTTCCGGGAGATCACCATGGGGCACACGATCGTGATGGGCCGGCGCACATG GGATTCGCTGCCGGCTAAAGTCCGGCCGCTGCCCGGCCGGCGAAATGTCGTACTGAGCCGCCAAGCTGAC TTTATGGCCAGCGGGGCTGAGGTTGTCGGTTCACTCGAGGAGGCGCTGACCAGCCCGGAGACGTGGGTGA TCGGAGGCGGACAAGTCTATGCGCTGGCGCTGCCGTACGCGACCAGATGTGAGGTTACCGAGGTCGACAT CGGCCTGCCGCGCGAAGCCGGTGACGCGCTGGCCCCCGTGCTGGACGAGACATGGCGGGGCGAGACGGGG GAGTGGCGCTTCAGCCGGTCCGGGTTGCGGTACCGGTTGTACAGCTACCACCGCTCATGA code 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):**