TargetSp15+-+CAI-1autoinducer+ synthase+(Vibrio+Cholerae)


 * *Target (protein/gene name): ** CAI-1 autoinducer synthase/cqsA

Cholera is an acute, diarrheal illness caused by infection of the intestine with the bacterium //Vibrio cholerae//. G lobally, cholera cases have increased steadily since 2005 and the disease still occurs in many places including Africa, Southeast Asia, and Haiti**. ** An estimated 3-5 million cases and over 100,000 deaths occur each year around the world. The infection is often mild or without symptoms, but can sometimes be severe. Approximately one in 10 (5-10%) infected persons will have severe disease characterized by profuse watery diarrhea, vomiting, and leg cramps. In these people, rapid loss of body fluids leads to dehydration and shock. Without treatment, death can occur within hours.
 * *NCBI Gene # or RefSeq#: **
 * *Protein ID (NP or XP #) or Wolbachia#: ** 243277
 * *Organism (including strain): **// Vibrio cholerae serotype O1 (strain ATCC 39315 / El Tor Inaba N16961) //
 * Etiologic Risk Group (see link below): ** Risk Group lv 2 (moderate individual risk, limited community risk)
 * */ Disease Information (sort of like the Intro to your Mini __Research Write__ up): **
 * Link to TDR Targets page (if present): **
 * Link to Gene Database page (NCBI, EuPath databases -e.g. TryTryp, PlasmoDB, etc - or PATRIC, etc.): **
 * [] ` **
 * Essentiality of this protein: ** Quorum sensing one another and to regulate a wide variety of physiological activities

[]

Is it a monomer or multimer as biological unit ** ? (make prediction at ** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ): Monomer
 * Complex of proteins?: **
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **


 * *EC#: ** 2.3.1.194
 * Link to BRENDA EC# page: ** []
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic



= [|**http://www.sigmaaldrich.com/catalog/search?term=CAI-1+autoinducer+synthase%2FcqsA&interface=All&N=0+220003052&mode=partialmax&lang=en&region=US&focus=site**] = = [|**http://www.sciencedirect.com/science/article/pii/S002228360900881X**] =
 * Enzyme Assay information (spectrophotometric, coupled assay ?, reagents): **
 * -- link to Sigma (or other company ) page for assay (see Sigma links below) **
 * -- -or link (or citation) to paper that contains assay information **
 * -- links to assay reagents (substrates) pages. **
 * --- List cost and quantity of substrate reagents, supplier, and catalog # **

-- PDB # or closest PDB entry if using homology model: -- For Homology Model option: Show pairwise alignment of your BLASTP search in NCBI against the PDB Query Coverage: Max % Identities: % Positives Chain used for homology:
 * Structure (PDB or Homology model) **

// Escherichia coli // overexpressing //V. harveyi cqsA// (WN1327) was grown overnight in LB with kanamycin at 30°C with shaking. The culture was diluted 1000-fold in M9 minimal salts medium (Sigma) supplemented with 0.05% (w/v) leucine, 2 mM MgCl2, 0.1 mM CaCl2, 0.01% (w/v) thiamine and 0.4% (w/v) glucose with 10 mg l−1 kanamycin. The culture was incubated overnight at 30°C with shaking. Cells were removed by centrifugation and the cleared fluid was extracted by DCM. The extract was carefully concentrated without heating to a small volume (5 ml) in a Rotovap. The concentrated extract was fractionated using HPLC (see //Supporting information//). Each HPLC fraction was assayed in triplicate for //V. harveyi// CqsS agonist activity using the reporter strain JMH626 as described above. Fractions containing activity were further characterized.
 * Current Inhibitors: ** neoglycoprotein
 * Expression Information (has it been expressed in bacterial cells): **// Escherichia coli //
 * Purification Method : **
 * Image of protein (PyMol with features delineated and shown separately): **



MNKPQLPDFI QNKIDHYIEN YFDINKNGKH LVLGKQASPD DIILQSNDYL
 * [] **
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **

ALANHPLIKA RLAKSLLEEQ QSLFMSASFL QNDYDKPMIE KRLAKFTGFD

ECLLSQSGWN ANVGLLQTIC QPNTNVYIDF FAHMSLWEGA RYANAQAHPF

MHNNCDHLRM LIQRHGPGII VVDSIYSTLG TIAPLAELVN ISKEFGCALL

VDESHSLGTH GPNGAGLLAE LGLTREVHFM TASLAKTFAY RAGAIWCNNE

VNRCVPFISY PAIFSSTLLP YEAAGLETTL EIIESADNRR QHLDRMARKL

RIGLSQLGLT IRSESQIIGL ETGDERNTEK VRDYLESNGV FGSVFCRPAT

389 43593.6  36370
 * SKNKNIIRLS LNSDVNDEQI AKIIEVCSDA VNYGDFYFR **
 * *length of your protein in Amino Acids: **
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: **
 * Molar Extinction coefficient of your protein at 280 nm wavelength: **
 * 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): **
 * *GC% Content for gene: **
 * *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) -- 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)

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