User:Nkuldell:M13 redesign: Difference between revisions

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[[M13 redesign, expt'l details]]
==Due Diligence==
==Due Diligence==
===Life Cycle of the Phage===
===M13 phage proteins===
===M13 phage proteins===
NCBI link to seq: [[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=56713234]] <br>
[[Image:Macintosh HD-Users-nkuldell-Desktop-M13K07 plasmid-phageproteins.jpeg|thumb|left|500px| phage proteins]]
Source: Phage Display of Peptides and Proteins: a laboratory manual. Kay, Winter, McCafferty. Ch 1. [[http://www.amazon.com/Phage-Display-Peptides-Proteins-Laboratory/dp/0124023800]]<br>
[[Image:Macintosh HD-Users-nkuldell-Desktop-M13K07 plasmid-phageparticle.jpeg|thumb|left| phage particle cartoon]] <br> <br> <br>
*NCBI link to seq: [[http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?db=nucleotide&val=56713234]] <br>
*Source: Phage Display of Peptides and Proteins: a laboratory manual. Kay, Winter, McCafferty. Ch 1. [[http://www.amazon.com/Phage-Display-Peptides-Proteins-Laboratory/dp/0124023800]]<br>
{| border="1"
{| border="1"
! protein
! protein
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| translation reinitiation inside gI  
| translation reinitiation inside gI  
|}
|}
===Vectors for M13 protein expression===
 
*M13KE [[http://www.westburg.nl/pages/TechnicalResources/Vectors.htm]]
===Life Cycle of the Phage===
*pUC118 [[http://seq.yeastgenome.org/vectordb/vector_descrip/PUC118.html]] or 119 [[http://seq.yeastgenome.org/vectordb/vector_descrip/PUC119.html]]
[[Image:Macintosh HD-Users-nkuldell-Desktop-M13K07 plasmid-M13lifecycle.jpeg|thumb|left|350px| cartoon depiction for life cycle of M13]]
*M13K07 (helper phage)[[http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/sequences/m13ko7.txt |sequence]][[http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/sequences/m13ko7.txt]]
[[Image:Macintosh HD-Users-nkuldell-Desktop-M13K07 plasmid-lifecycle key.jpeg|thumb|left|500px| key]]
==Makeover ideas==
==Makeover ideas==
===phage or phage coat rewrite===  
===phage or phage coat rewrite===  
===separate 8, 9, 3===
===separate 8, 9, 3===
===two copies of 8===
===two copies of 8===

Latest revision as of 10:58, 26 October 2006

M13 redesign, expt'l details

Due Diligence

M13 phage proteins

phage proteins
phage particle cartoon




  • NCBI link to seq: [[1]]
  • Source: Phage Display of Peptides and Proteins: a laboratory manual. Kay, Winter, McCafferty. Ch 1. [[2]]
protein aa (mature) function notes
I 348 assembly includes gXI within, the two form structure in cytoplasmic membrane for phage maturation
II 410 replication of DNA + strand includes gX within
III 406 phage tail protein (5 copies) tag at N-term
IV 405 assembly organizes to pore in OM
V 87 binds ssDNA recognizes DNA PS
VI 112 phage tail protein (5 copies) packages with pIII
VII 33 phage head protein (5 copies) packages with p9
VIII 50 phage coat protein (2700 copies) seg seq cleaved, matured form at aa 23 = clones to N-term, "Petrenko" site [[3]]
IX 32 phage head protein (5 copies) stop codon overlaps start of gVIII
X 111 DNA replication internal reinitiation inside gII
XI 108 assembly translation reinitiation inside gI

Life Cycle of the Phage

cartoon depiction for life cycle of M13
key

Makeover ideas

phage or phage coat rewrite

separate 8, 9, 3

two copies of 8