TargetSp15+-+Penicillin-binding+protein+2x+(Streptococcus+Pneumoniae)


 * *Target (protein/gene name): ** Penicillin-binding protein 2x
 * *NCBI Gene # or RefSeq#: ** [|170187]
 * *Protein ID (NP or XP #) or Wolbachia#: ** NC_008533.1
 * *Organism (including strain): **// S. pneumoniae (strain ATCC BAA-334 / TIGR4) //

// Streptococcus pneumoniae (S. pneumoniae) // are lancet-shaped, gram-positive, facultative anaerobic bacteria with over 90 known serotypes. Most //S. pneumoniae// serotypes have been shown to cause disease, but only a minority of serotypes produce the majority of pneumococcal infections. Pneumococci are common inhabitants of the respiratory tract and may be isolated from the nasopharynx of 5-70% of adults, depending on the population and setting. Only 5-10% of adults without children are carriers. In schools and orphanages, 25-50% of students and residents may be carriers. On military installations, as many as 50-60% of service personnel may be carriers. The duration of carriage varies and is generally longer in children than adults. In addition, the relationship of carriage to the development of natural immunity is poorly understood.
 * Etiologic Risk Group (see link below): ** Risk Group lv 2 (moderate individual risk, low 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: ** PBP2x constitute primary resistance determinants and confer low-level β-lactam resistance
 * (make prediction at ** @http://www.ebi.ac.uk/msd-srv/prot_int/pistart.html ): Mononer
 * Complex of proteins?: **
 * Druggable Target (list number or cite evidence from a paper/database showing druggable in another organism): **


 * *EC#: **** 2.1.2.2 **
 * Link to BRENDA EC# page: **
 * [] **
 * -- ** Show screenshot of BRENDA enzyme mechanism schematic

[[image:Screen Shot 2015-04-21 at 3.07.04 AM.jpg width="800" height="294" caption=" Reaction catalyzed by phosphoribosylglycinamide formyltransferase (2.1.2.2)"]]


 * 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) **


 * Current Inhibitors: **** Penicillin **
 * Expression Information (has it been expressed in bacterial cells): Yes **

DNA fragments were purified with the QIAEX II gel extraction kit (Qiagen, Hilden, Germany) according to the manufacturer’s recommendations. PCR-derived DNA fragments were cloned into the PCR II vector (Invitrogen). DNA sequences were determined for at least two clones. Cell walls were washed in H2O–1 M NaCl to remove the SDS and then purified by incubation with 200 μg of pronase/ml for 16 h, which was followed by the addition of 200 μg of trypsin/ml for 16 h. The mixture was then washed three times with 1% SDS, 8 M LiCl, and 100 mM EDTA and then was washed at least twice with H2O
 * Purification Method : **
 * Image of protein (PyMol with features delineated and shown separately): **

http://www.rcsb.org/pdb/explore/explore.do?structureId=4S1N

MKWTKRVIRY ATKNRKSPAE NRRRVGKSLS LLSVFVFAIF LVNFAVIIGT GTRFGTDLAK EAKKVHQTTR TVPAKRGTIY DRNGVPIAED ATSYNVYAVI DENYKSATGK ILYVEKTQFN KVAEVFHKYL DMEESYVREQ LSQPNLKQVS FGAKGNGITY ANMMSIKKEL EAAEVKGIDF TTSPNRSYPN GQFASSFIGL AQLHENEDGS KSLLGTSGME SSLNSILAGT DGIITYEKDR LGNIVPGTEQ VSQRTMDGKD VYTTISSPLQ SFMETQMDAF QEKVKGKYMT ATLVSAKTGE ILATTQRPTF DADTKEGITE DFVWRDILYQ SNYEPGSTMK VMMLAAAIDN NTFPGGEVFN SSELKIADAT IRDWDVNEGL TGGRTMTFSQ GFAHSSNVGM TLLEQKMGDA TWLDYLNRFK FGVPTRFGLT DEYAGQLPAD NIVNIAQSSF GQGISVTQTQ MIRAFTAIAN DGVMLEPKFI SAIYDPNDQT ARKSQKEIVG NPVSKDAASL TRTNMVLVGT DPVYGTMYNH STGKPTVTVP GQNVALKSGT AQIADEKNGG YLVGLTDYIF SAVSMSPAEN PDFILYVTVQ QPEHYSGIQL GEFANPILER ASAMKDSLNL QTTAKALEQV SQQSPYPMPS VKDISPGDLA EELRRNLVQP IVVGTGTKIK NSSAEEGKNL APNQQVLILS DKAEEVPDMY GWTKETAETL AKWLNIELEF QGSGSTVQKQ DVRANTAIKD IKKITLTLGD
 * *Amino Acid Sequence (paste as text only - not as screenshot or as 'code'): **
 * *length of your protein in Amino Acids: ** 750
 * Molecular Weight of your protein in kiloDaltons using the [|Expasy ProtParam] website: ** 82313.0
 * Molar Extinction coefficient of your protein at 280 nm wavelength: ** 71740
 * 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)


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