Target+-+Protein+Tyrosine+Phosphatase+(Listeria+monocytogenes)

** Target (protein / gene name):   **  Protein Tyrosine Phosphatase - lmo1800

 * * NCBI Gene # or RefSeq #:  **   Project:  [|61583] ,    NC_003210.1, Gene ID: 985934


 * * Protein ID (NP or XP #) or Wolbachia #:  **  NP_465325.1 (use De La O's Oligos for ordering - Dr. B 090313)


 * * Organism:  **  Listeria monocytogenes


 * Etiologic Risk Group (see link below):  **  Risk Group 2 - Bacterial agents including Chlaymidia


 * * Background / Disease Information:  **  The Most Deadly Born food pathogen. Induces muscle aches, fever and diarrhea. Main route of infection is through uncooked and unwashed food.

[]
 * Essentiality of this protein:  **  Protein required to induce host to engulf bacteria. Upon entry to the cytosol bacteria can reproduce.


 * Complex of enzymes:  **  No


 * * EC #:  **  3.1.3.48


 * Link to BRENDA EC # page:  **[|http://brenda-enzymes.org/php/result_flat.php4?ecno=3.1.3.48]


 * Enzyme Assay information (spherophophometrics, coupled assay, reagents):  **


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

[| http://www.sigmaaldrich.com/catalog/search?interface=All&term=3.1.3.48&lang=en®ion=US&focuseddocuments&N = 0 + 220003049 + 219853269 + 219853286 & mode = match% 20partialmax] ?????


 * - link to paper that contains information Assay:  **

[| http://www.brenda-enzymes.org/literature/lit.php4?e=3.1.3.48&r=663621]


 * - List of cost and QUANTITY substrate reagents and Supplier  **

[|https://www.atcc.org/ATCCAdvancedCatalogSearch/ProductDetails/tabid/452/Default.aspx?ATCCNum=BAA-679D-5&Template=Bacteria]


 * Structure Available (PDB or Homology model)  ** [| http://www.rcsb.org/pdb /explore/explore.do?structureId=1YWF]**  - QUERY Coverage (Not exposed to direct match): 84%  ****  E value - 8 x 10 ^ -22  ****  Druggable Target (see databases for this): Yes  ****  Current Inhibitors:  ** None,  [|http: / / www.bindingdb.org/bind/ByMonomersTarget.jsp]**  Expression Information:  ****  Purification Method:  ****  PyMol or other:  ** Homology model with M. tuberculosis: Blastp gives 47% positives and 83 % query query. **  * Amino Acid Sequence:  **

http://www.rcsb.org/pdb/explore/explore.do?structureId=1YWF

> gi | 16803840 | ref | NP_465325.1 | lmo1800 hypothetical protein [Listeria monocytogenes EGD-e] MKNWVKVTGAGVLSATLLLGGCGAQSEEKAEANVKTEQTLKPGSQIKLEGAVNVRDLGGYKTTDGLTIKPHKLIRSAEL ANLSDSDKKKLVNTYDLSHIVDFRTSSEVATKPDPKLTDVDYTHDSVMKDNGTSTSTQDLTASLAKMDNPETFLINANKS FITDETSIQAYKDFFDILLANQDGSVLWHCTAGKDRAGFGTALVLSALGVDKNTVIDDYMLSNKYRADENKKAIEAVAAKT DNKKVIDGMTAVMEVRESYINAAFDEINAKYGSMDNFLKEKLGLTDAKKEQLKKAYLY


 * Molecular Weight of protein in your kiloDaltons using the [|Expasy ProtParam]  website   **
 * Molar Extinction coefficient of your protein at 280 nm wavelength:  **

> gi | 16802048: c1873620- 1872724 Listeria monocytogenes EGD-e, complete genome   ATGAAAAATTGGGTAAAAGTAACAGGAGCAGGGGTACTAAGCGCGACTTTACTATTAGGTGGATGCGGGG CGCAGTCAGAAGAAAAAGCAGAAGCAAATGTTAAAACCGAGCAAACACTCAAACCAGGAAGCCAAATTAA ATTAGAAGGCGCTGTAAATGTCCGGGACTTAGGCGGATACAAAACAACGGATGGACTAACCATTAAGCCA CATAAACTCATTAGAAGTGCCGAACTCGCTAACTTAAGTGATTCGGATAAAAAGAAACTCGTAAATACAT ATGATTTGTCTCATATAGTTGATTTCCGAACAAGTTCAGAAGTCGCAACGAAACCAGATCCGAAACTCAC AGATGTAGACTATACGCACGATTCTGTGATGAAAGATAATGGAACATCTACAAGCACACAAGATTTAACT GCTAGCCTAGCCAAAATGGATAATCCAGAGACATTTCTCATTAATGCTAATAAGAGCTTTATTACAGATG AAACTTCGATACAAGCCTATAAAGATTTTTTTGATATACTACTAGCAAACCAAGATGGTTCTGTTCTTTG GCACTGTACAGCTGGAAAAGACCGAGCTGGATTTGGAACCGCCCTCGTTCTTTCAGCGTTAGGTGTGGAT AAAAACACGGTCATTGACGATTATATGCTGTCCAATAAATATCGTGCTGACGAAAATAAAAAAGCAATTG AAGCCGTTGCAGCAAAAACAGATAATAAAAAAGTGATTGATGGAATGACAGCCGTAATGGAAGTTCGTGA ATCTTATATCAATGCAGCCTTCGATGAAATTAATGCAAAATATGGTTCGATGGACAACTTTTTAAAAGAAAAACTCGGACTAACCGATGCTAATAACAACAACTAAAAAAAGCATATCTTTATTAA
 * * CDS Gene sequence:  **
 * Partial  **


 * * GC% Content for gene: 37.9%  **

ATGAAAAACTGGGTTAAAGTTACCGGTGCGGGCGTTCTCTCTGCTACTCTGCTGCTCGGT GGTTGTGGTGCGCAGTCTGAGGAAAAAGCGGAAGCAAACGTCAAGACCGAACAGACCCTG AAGCCGGGTAGCCAGATCAAACTGGAAGGTGCCGTTAATGTTCGCGACCTGGGTGGTTAC AAGACTACCGATGGTCTGACCATCAAACCGCACAAACTGATCCGCTCTGCGGAACTCGCA AACCTCTCTGACTCTGACAAAAAGAAGCTGGTTAACACCTACGACCTGTCTCACATCGTT GACTTCCGCACGTCTTCTGAAGTTGCGACCAAACCGGACCCGAAACTGACCGACGTGGAC TACACCCACGACTCTGTTATGAAAGACAACGGTACGTCTACCTCTACCCAGGACCTGACG GCGTCTCTGGCCAAGATGGACAACCCGGAAACCTTCCTGATTAACGCGAATAAATCCTTC ATTACCGACGAGACCTCTATCCAGGCGTACAAAGACTTCTTCGACATCCTGCTGGCGAAC CAGGACGGTTCCGTTCTGTGGCACTGCACCGCCGGTAAAGACCGTGCCGGTTTCGGCACC GCGCTGGTTCTGTCTGCGCTGGGTGTTGATAAAAACACCGTTATCGACGACTACATGCTG TCTAACAAATACCGTGCTGACGAAAATAAGAAAGCGATCGAAGCGGTAGCGGCGAAAACC GACAACAAAAAAGTGATCGACGGTATGACGGCAGTTATGGAAGTTCGTGAATCTTACATC AACGCGGCGTTCGATGAGATCAATGCGAAATATGGTTCTATGGATAACTTCCTCAAAGAAAAGCTGGGCCTCACTGATGCGAAGAAAGAGCAGCTGAAAAAAGCATACCTGTACTAA
 * * CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol:  **


 * *% GC content for Gene (codon optimized): 50.9%  **


 * primer design results for pNIC-Bsa4 cloning (list of all of your seqeunces ~ 40 nt long primers)  **

 Oligos sequence

Lmon_PTP_1 || ATGAAGAATTGGGTTAAAGTTACCGGTGCAGGTGTTCTGTCCGCGACCCTGCTGCT || || Lmon_PTP_2 || ACGTTCGCTTCCGCTTTTCCTCAGACTGCGCACCACAACCACCCAGCAGCAGGGTCGCG || || Lmon_PTP_3 || WHERE YOU SHOULD BE A BETWEEN YOUCCCCECCAACCGGTCTCTCAAATCAAACT || || Lmon_PTP_4 || TTATAACCGCCGAGATCGCGCACGTTAACCGCACCTTCCAGTTTGATTTGAGAACCCGGT || || Lmon_PTP_5 || GCGATCTCGGCGGTTATAAGACTACCGACGGTCTCACGATCAAACCGCATAAACTGATCC || || Lmon_PTP_6 || TCTTATCAGAGTCAGACAGATTCGCCAGTTCCGCAGAACGGATCAGTTTGGGTTTGA || || Lmon_PTP_7 || CGAATCTGTCTGACTCTGATAAGAAAAAACTGGTTAACACCTACGACCTGTCTCACATCG || || Lmon_PTP_8 || TCCGGTTTGGTCGCAACTTCAGAAGAGGTGCGAAAGTCAACGATGTGAGACAGGTCGTAG || || Lmon_PTP_9 || GTTGCGACCAAACCGGACCCAAAGCTGACCGACGTGGATTACACCCACGACTCTGTTATG || || Lmon_PTP_10 || GGTCAGGTCTTGGGTGGAGGTAGAGGTACCGTTGTCTTTCATAACAGAGTCGTGGGTGTA || || Lmon_PTP_11 || TCCACCCAAGACCTGACCGCGTCTCTCGCGAAGATGGATAACCCGGAAACCTTCCTGATC || || Lmon_PTP_12 || TGGATAGAGGTTTCATCGGTGATAAAAGATTTGTTCGCATTGATCAGGAAGGTTTCCGGG || || Lmon_PTP_13 || CACCGATGAAACCTCTATCCAGGCGTACAAAGACTTCTTCGACATCCTGCTCGCAAACCA || || Lmon_PTP_14 || CGGTCTTTACCCGCAGTGCAGTGCCACAGGACAGAGCCGTCTTGGTTTGCGAGCAGGATG || || Lmon_PTP_15 || CACTGCGGTAAAGACCGTGCTGGTTTCGGCACCGCGCTGGTCTCTCTGCACTCGGTGT || || Lmon_PTP_16 || TGTTAGACAGCATGTAGTCGTCAATAACGGTGTTTTTGTCCACACCGAGTGCAGAGAGAA || || Lmon_PTP_17 || ACGACTACATGCTGTCTAACAAATACCGTGCGGACGAAAACAAAAAGGCGATCGAAGCGG || || Lmon_PTP_18 || GTCATACCGTCGATAACTTTCTTGTTGTCGGTTTTCGCCGCAACCGTTTCGATCGCCTTT || || Lmon_PTP_19 || CAAGAAAGTTATCGACGGTATGACCGCAGTTATGGAAGTTCGTGAATCTTACATTAACGC || || Lmon_PTP_20 || ATAGAACCGTATTTTGCGTTGATTTCGTCGAACGCCGCGTTAATGTAAGATTCACGAACT || || Lmon_PTP_21 || ATCAACGCAAAATACGGTTCTATGGACAACTTCCTCAAGGAGAAACTGGGTCTGACGGAC || || Lmon_PTP_22 || TTAGTACAGGTACGCTTTCTTCAGCTGTTCTTTTTCTCGCGTCCGTCAGACCCAGTTTC ||

Primers LmPTP-for imc   TACTTCCAATCC  __  ATG  __ __   AAGAACTGGGTTAAAGTT   __ LmPTP-rev imc   AAAAGGCATACCTGTACTAA     CAGTAAAGGTGGATA  <span style="color: #0070c0; font-family: 'Times New Roman',serif; font-size: 12pt;"> Primers - C. Burns - Underlined is partial nucleotide sequence - middle black are start and stop codons Forward - TACTTCCAATCC ATG __ ACCCTCGTTCTGCGTTAT __ Reverse - TATCCACCTTTACTG TTA __ GTGTTCGAGGTCCGC __ - Ask a mentor, Dr. B, or a fellow researcher -how to link a GDocs file if you are not sure how to. []
 * (    link to DNA Works output text file -   **   that should be saved in your Google Docs folder after you did the primary design protocol)

TMpred Output: