TargetSp17-+type+II+site-specific+deoxyribonuclease(Escherichia+coli)

Escherichia Coli are bacteria that live in the intestines and are usually harmless but some of them can be pathogenic. They can cause diarrhea or infection outside of the intestinal tract. These types of bacteria are transmitted through contaminated water, food, or contact with another person or animal. The six types of E.Coli that cause this disease are Shiga toxin producing E.coli (STEC), Enterotoxigenic E. coli (ETEC), Enteropathogenic E. coli (EPEC), Enteroaggregative E. coli (EAEC), Enteroinvasive E. coli (EIEC) and Diffusely adherent E. coli (DAEC). Unfortunately, there is not a cure for E.Coli infections, it has to run its course and eventually get out of the body. To prevent from contracting en E.Coli infection maintain hands clean, cook meat thoroughly, wash vegetables, store foods separately, and get pasteurized drinks. Links:
 * Target (protein):** Type II site-specific deoxyribonuclease
 * E.C Number:** 3.1.21.4
 * Protein ID:** WP_000781649.1
 * PDB ID:** 1STX
 * Organism:** Escherichia coli
 * Etiologic Risk Group (see link below): N/a **
 * Disease Information:**
 * 1) []
 * 2) []
 * 3) [|https://www.patricbrc.org/view/Feature/RefSeq.356829.3.JGZU01000001.CDS.4223.5182.fw]


 * Link to TDR Targets page (if present): N/A **

The Type II site-specific deoxyribonuclease is an enzyme that helps catalyze en donucleolytic cleavage of DNA to give specific double-stranded fragments with terminal 5'-phosphates. Links:
 * Essentiality of Type II site-specific deoxyribonuclease:**
 * 1) []

**Druggable Target:** Not available


 * Link to BRENDA: **[]


 * Enzyme Assay Information:** http://www.sciencedirect.com/science/article/pii/S002228360301297X

Eco RV assay of DNA using N6-methyladenine

[]

Relative activity in PCR mix (Taq DNA Polymerase buffer) is 10%. The PCR mix contained λDNA, primers, 10 mM Tris-HCl (pH 8.3, 20 °C), 50 mM KCl, 1.5 mM MgCl2, 200 μM dNTPs, 2.5 U Taq DNA polymerase. The mix was subjected to 25 amplification cycles. Activity in reaction buffer of Pwo SuperYield DNA Polymerase PCR Mix is 25%. When supplemented with GC-RICH Solution activity remains at 25%. The Eco RV assay that contains all of these materials in it is worth $62.
 * Reagents and Materials:**


 * Structure (PDB) **
 * Sequence ID: ** WP_000781649.1

Show pairwise alignment of your BLASTP search in NCBI:

**Query Coverage:** 100% **Max % Identities:** 100% **% Positives:** 100% **Chain used for homology:** A


 * Inhibitors:** EDTA

The target has been expressed in E.Coli. It is expressed predominatley in bacteria because it is a restriction modification system that helps protect the bacteria from foreign DNA that tries to inhibit it.
 * Expression Information:**
 * Link:**
 * 1) []


 * Purification Method:** Heparin-Sepharose column chromatography


 * Image of Protein:**

Figure 1: Structure of the K38A mutant of EcoRV bound to cognate DNA and Mn2

code 1__0__        2__0__         3__0__         4__0__         5__0__         6__0__ MNNQNDYIEA AQQIIASLGL PRAQQNERSA LCLLALLNLT PGKAWADAEN PLVGITPIMN
 * Amino Acid Sequence (paste as text only - not as screenshot or as 'code'):**

7__0__        8__0__         9__0__        10__0__        11__0__        12__0__ WVREHYGKVY APNTRETFRR QSMHQFCAAG VALYNPDKPD RPVNSPKAVY QIEPAALSML

13__0__       14__0__        15__0__        16__0__        17__0__        18__0__ RTFGSPAWHD SLATYLAERE TLVTRYAKER EQNRIPVEIA AGQQITLSPG EHSELIRAII

19__0__       20__0__        21__0__        22__0__        23__0__        24__0__ EDFAPRFAPG SVLVYAGDTG EKWGYFDAPL LAGLGVDVDS HGKMPDVVLH FTAKNWLLLV

25__0__       26__0__        27__0__        28__0__        29__0__        30__0__ ESVTSHGPVD GKRHAELARL FAGSTAGLVY VTAFPNRSIM GRYLGEIAWE TEVWVADAPS

31__0__ HLIHFNGVRF LGPYSTE code **Molecular Weight** **of your protein in kiloDaltons using the [|Expasy ProtParam] website:** 35064.82 KDa
 * length of your protein in Amino Acids:** 317
 * Molar Extinction coefficient of your protein at 280 nm wavelength:** 56505
 * TMpred graph Image (@http://www.ch.embnet.org/software/TMPRED_form.html). Input your amino acid sequence to it.**

Figure 2: TMPred output graph for Type II site-specific deoxyribonuclease code NC_017527.1:757901-761041 Methanosaeta harundinacea 6Ac, complete genome GTGGCCGTTCCGGAGAAGATCCTTGAGTTGGTTGAAAAATTCGAATTGGTCATTGATAAATATAAATCTA GTTATAAAGAAGAAGAAATTAAAATTGAATTTATAAATCCTTTTTTTGAAGCTCTAGGCTGGGATGTCCG AAATAAGAGCAATAACCCCCCAGATCAGAGAGAGGTCAGGTTTGAAGATACCTTGAGGTCAGAGGGGGAG ACCCAGAGGCCCGATTATAGCTTCTGGCTTGACAGGGAGAGGAAGTTCTTTCTGGACGCCAAAAAACCTT CAGTGAATATTGAAGGAGGAATAATCCCAGCATTCCAGATAAAGCGTTATGCATACTCTTCTACCCTGCC GCTTGCCATTGTAACCGACTTTGAAGAGTTTGCCGTTTACGATTGCAGGATCAAACCGTTCAAATTTGAT AAGCCACAAAAGTCTAGATTATTATTAATAAAATATTATGAATATGAAAAGAGATGGGACGAGATCGCCT CGTTGTTCTCCAAAGAGGCCGTTCAGAGAGGCTCCCTCGGCTCCATCCCTCCGCCAAAAGGAAAAAAAGT CGACGACGCTCTTCTGGAGGACATCTCTGATTGGCGAAATGCTCTGGCGGCGAGCATCACTAAGATGAAT CCAGAGATCTCTCAGAAGAGCCTGAACTATGCGGTCCAGATGACGATCGACAGGATTCTATTTCTAAGGA TCTGTGAGGATCGGTCTATTGAGGACAGCGAGCGGTTGAAGGGGCTTCTTGATGGGGATGATGTATATAA GAGGCTGTTTGAACTCTTCGAAGAGGCCGATAAGAGGTACAATTCGGGGCTGTTCCACTTCCATCCTGAG GAGGAGAGGAACAACTTCGACGCCATCACCCCGTCGATAAAGATCGAGGACAATGTTCTTAAGAATATAA TTCAAAGCTTATACTATCCTGACAGCCCCTACGTCTTCTCTGAGATCCCTGCCGATATTCTTGGTCACGT TTACGAGCAGTTCCTGGGGAAGGTCATCCAGCTGGACGAGAGCCACAAAGTGAGCGTGGTGGAGAAGCCT GAGGTGAGGAAGGCAGGAGGCGTTTATTATACTCCATCATATATTGTAGATTATATCGTTAAAAACACCG TCGGCAAGCTCTTGGAGGGGAAGACCCCCAAGGAGGCCTCTGACCTTCGCATCCTCGATCCCGCCTGCGG TTCGGGGTCGTTTCTGATAGTGGCCTATCAGCACCTCTTGGATTGGCACCTCGACTGGTACAAGGCCCAT CTCGTCCCTCTCTTGGATGCTGGAGAGAAGCCGACATCAAACGGGGTGCAAAATCTCCTCCCGTCTCCGA TCGAGAGGCCAGGGAAGAACGCCAGGTCCCGATCGAAGAAGATCTGGGAGAGGGCTTCGGTGGCGACCCT CCCCATCGAGGAGAGGGCGGGAGAATGGCATCTGAAGACCGCCGAGAGGAAGAGGATCCTCCTCAACAAC ATCTATGGCGTGGATATAGACACTCAGGCCGTGGAGGTGACAAAGCTCTCCCTCCTCCTCAAGGTCTTGG AAGGAGAAAATAAGGCCACCATCACCGATCTCAACATGTATTCAGATGAGAGGGCTCTGCCAGATCTTGG AAATAATATAAAATGTGGCAATTCCCTAATCGGTTGGGATATCCTTGAAGATAAGCCCGATCTCTCCCAG GAAGAGCTGGAGAGGATCAATCCATTTATATGGGAGGATGAATTTAAAGAGGTCTTCGCCAAAGGCGGTT TTGATGCGGTAATTGGAAATCCGCCTTATATTAGAATTCAAATGATGAAGGAATGGGCTCCTGTAGAGGT GGAATTTTACAAAGAGCATTACACGGTCGCCAAGAAAGGGAACTATGACGTTTATGTCGTATTCGTCGAA AAGGGATTGGCCCTCCTGAACGAAAGCGGTCGTTTAGGTTTTATCCTACCAAATAAATTTTTTAATGCAC AATATGGTTCACCGTTGCGTAGTTTATTATCAAATGGAAAAAATTTAGACGAAATTGTTAATTTTGGGCA TGGACAAATATTCTGTGGCGTAACTACCTATACATGTCTGATCTTTCTGAATAAAATGCCAAAAGATTCA TTCAAATATACATATGTCGAAGACTTGAATTCATGGAAAGAGGACGCATTATCAATAACGGGTGAAATTC AAGCTAAGAATGCAGCATGTGCTGAATGGAATTTTATTGTTGGAAAAGGTTCAGATCTTTTTGATCGTTT GAGTAAGATGCCTGTGAAGTTGGGAGATGTGGCTGAAAGGGTGTTTCAAGGGATCATTCCTGGAATGGAT GCAGTTTATTCGATAAAATTGATCGGGGAAGATGATATTGTAATATGTTATTCGCGGGCTTTATCTAAAG ATATTCAGATAGAAAGTAAAATTCTTCGTAAAATCGCCAGCGGTTCTCAGGTGAAGGCTTATACAATTGT AGATGATAAATCAAGAGTTATCTATCCTTATCACTTTGAGGGAGATAATTGTGTTTTAATTCTGCCAGAT AAGATGAAACAGGATTTTCCTTTGGCTTATGATTATCTTCAAAAAACCAGAGATCTATTGGACAAAAGAG ATCGTGGCTCGGCTAAAGGAGCCCAATGGTATCGTTATATTAGAACCCAAAATATTGGATTACAAGCATA TCCTAAGTTGGCCATCCCTCGCCTCGTCCATCGCTTGTGTGCAGCCTATGACGCAAATGCAGATTTTTGT TTGGATAACGTGGATGTGGGTGGAGTGATTCTCTCTGAAAATAGCCCTGTTGGTTATATTTATATACTTG GCTTATTAAATAGCAAGCTACTCAACTATTATTTTAAAATTAATAGTGTTCATTTTCGAGGTGGTTTTTT CTCTGCTAATAGGCAATATATTGAACGTCTCCCCATTCGCACCATCGACTTCTCCGACCCCCAGGACGTC ACCCGCCACGACAAGATGGTGAGCCTCGTGGAGACGATGCTCGACCTGCACAAACAACTCCAGGAAGTCG GCACGCCACATGAAAAAACGCGCATCCAAAATCAGATCGGTTATACGGACAAGCAGATAGATGCCTTGGT CTACGAACTGTACGACCTCACAGAGGAGGAGATCGCCATCGTGGAAGAGAGCACAAAATAG code
 * CDS Gene Sequence (paste as text only):**
 * GC% Content for gene:** 7.886435%
 * CDS Gene Sequence (codon optimized) - copy from output of Primer Design Protocol (paste as text only): N/a**
 * GC% Content for gene (codon optimized): N/a**