Target-Histidinol+Phosphatase+(Escherichia+coli+O157+H7)

= Target-Histidinol Phosphatase = The bacterium Escherichia coli O157:H7 is a worldwide threat to public health and has been implicated in many outbreaks of haemorrhagic colitis. In addition to causing diarrhea, Escherichia coli O157:H7 infection can lead to hemolytic-uremic syndrome (HUS), a severe disease characterized by hemolysis and renal failure. The severity of disease, the lack of effective treatment and the potential for large-scale outbreaks from contaminated food supplies have propelled intensive research on the pathogenesis and detection of E. coli O157:H7. 1,387 new genes encoded in strain-specific clusters of diverse sizes were found in O157:H7. These include candidate virulence factors, alternative metabolic capacities, several prophages and other new functions--all of which could be targets for surveillance. [|Genome Sequence Article] High-resolution phylogenomic approaches allow the dynamics of pathogenome evolution to be followed at a high level of phylogenetic accuracy and resolution. SNP discovery and study of genome architecture and prophage content identified numerous biomarkers to assess the extent of genetic diversity within a set of clinical and environmental strains. [|Genomic Anatomy Article] This information is not yet available and could not be found LB Media (Glucose)-$40.70 L3022-250G Indole-$64.30 I3408-100G Sigma Aldrich Supplier -- PDB # or closest PDB entry if using homology model: 2FPW -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB [|2FPU vs. 2FPW] Query Coverage: 176 letters Max % Identities: 163/163 (100%) % Positives: 163/163 (100%) Chain used for homology: B
 * Target (protein/gene name): Histidinol Phosphatase** (Histidine biosynthesis bifunctional protein hisB)
 * NCBI Gene # or RefSeq#:** GI:114793851
 * Protein ID (NP or XP #) or Wolbachia#:** NP_288527.2
 * Organism (including strain):** __Escherichia coli O157:H7__
 * Etiologic Risk Group (see link below):** __Risk Group 1__
 * Background/Disease Information (sort of like the Intro to your Mini Research Write up):**
 * Essentiality of this protein:** Histidinol Phosphatase is not essential.
 * Complex of proteins?:** No
 * Druggable Target:** There are no current druggable targets.
 * *EC#: ** 3.1.3.15
 * Link to BRENDA EC# page:** []
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): Spectrophotometric**
 * -- link to Sigma (or other company) page for assay or assay reagents (substrates)**
 * -- link (or citation) to paper that contains assay information**
 * (Page 4598)** []
 * -- List cost and quantity of substrate reagents and supplier**
 * Structure Available (PDB or Homology model)**

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 * Current Inhibitors:** HisB-N is active in the presence of Mg(2+), Mn(2+), Co(2+), or Zn(2+), but Ca(2+) has an inhibitory effect.[|Article Source]
 * Expression Information (has it been expressed in bacterial cells):** Yes, it has been expressed in E. Coli BL21 (DE3). [|Article Source]
 * Purification Method:** Purification Method is explained in the Cloning, Mutagenesis, and Purification section of the article below.
 * Image of protein (PyMol with features delineated and shown separately):**

MGSSHHHHHHGSSQKYLFIDRDGTLISEPPSDFQVDRFDKLAFEPGVIPQLLKLQKAGYKLVMITNQDGL GTQSFPQADFDGPHNLMMQIFTSQGVQFDEVLICPHLPADECDCRKPKVKLVERYLAEQAMDRANSYVIG DRATDIQLAENMGINGLRYDRETLNWPMIGEQLTRR [] Abs 0.1% (=1 g/l) 0.652, assuming all pairs of Cys residues form cystines [|NCBI Gene] >gi|16445223:2835746-2836813 Escherichia coli O157:H7 str. EDL933 chromosome, complete genome ATGAGTCAGAAGTATCTTTTTATCGATCGCGATGGAACCCTGATTAGCGAACCGCCGAGTGATTTTCAGG TGGATCGTTTTGACAAACTCGCCTTTGAACCGGGCGTGATCCCGCAACTGCTGAAGCTGCAAAAAGCGGG CTACAAACTGGTGATGATCACTAATCAGGATGGTCTGGGAACACAAAGTTTCCCGCAGGCGGATTTCGAT GGCCCGCACAACCTGATGATGCAGATATTCACYTCGCAAGGCGTACAGTTTGATGAAGTGCTGATTTGTC CGCACCTGCCCGCCGATGAGTGCGACTGCCGTAAACCGAAAGTAAAACTGGTGGAGCGTTATCTCGCTGA GCAAGCGATGGATCGCGCCAACAGTTATGTGATTGGCGATCGCGCGACCGATATTCAACTCGCTGAAAAC ATGGGCATTAATGGTTTACGCTACGACCGCGAAACCCTGAACTGGCCAATGATTGGCGAGCAACTCACCA GACGTGACCGTTACGCTCACGTAGTGCGTAATACCAAAGAGACGCAGATTGACGTTCAGGTGTGGCTGGA TCGTGAAGGTGGCAGCAAGATTAATACCGGCGTTGGCTTCTTTGATCACATGCTGGATCAGATCGCCACC CACGGCGGTTTCCGTATGGAAATCAACGTCAAAGGCGACCTCTATATCGACGATCACCACACCGTCGAAG ATACCGGCCTGGCGCTGGGCGAAGCGTTAAAAATTGCCCTCGGCGATAAACGCGGTATTTGCCGCTTTGG TTTTGTGCTGCCGATGGACGAATGCCTTGCCCGCTGCGCGCTGGATATCTCTGGTCGCCCGCACCTGGAA TATAAAGCCGAGTTTACCTACCAACGCGTGGGCGATCTCAGCACCGAGATGATCGAGCACTTCTTCCGTT CGCTCTCTTACACCATGGGCGTGACGCTCCACCTGAAAACCAAAGGTAAAAACGATCATCACCGTGTAGA GAGCCTGTTCAAAGCCTTTGGTCGGACCCTGCGCCAGGCCATCCGCGTGGAAGGCGATACCCTGCCCTCG TCGAAAGGAGTGCTGTAA
 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):**
 * length of your protein in Amino Acids:** 176
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website:** 20040.7
 * Molar Extinction coefficient of your protein at 280 nm wavelength:** 13075
 * TMpred graph Image ** ( @http://www.ch.embnet.org/software/TMPRED_form.html ). Input your amino acid sequence to it.[[image:http://www.ch.embnet.org/wwwtmp/.TMPRED.16903.7537.gif align="middle" caption="TMpred Graph Image of Amino Acid Sequence of Histidinol Phosphatase"]]
 * CDS Gene Sequence (paste as text only):**
 * GC% Content for gene: 53.18%**

-- 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 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)


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

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 * References:**