User:Nkuldell:M13 redesign

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Line 13: Line 13:
| 348
| 348
| assembly
| assembly
-
|  
+
| includes gXI within, the two form structure in cytoplasmic membrane for phage maturation
|-
|-
| II  
| II  
| 410
| 410
| replication of DNA + strand
| replication of DNA + strand
-
|  
+
| includes gX within
|-
|-
| III  
| III  
| 406
| 406
| phage tail protein (5 copies)
| phage tail protein (5 copies)
-
|  
+
| tag at N-term
|-
|-
| IV  
| IV  
| 405
| 405
| assembly
| assembly
-
|  
+
| organizes to pore in OM
|-
|-
| V  
| V  
| 87
| 87
| binds ssDNA
| binds ssDNA
-
|  
+
| recognizes DNA PS
|-
|-
| VI  
| VI  
| 112
| 112
| phage tail protein (5 copies)
| phage tail protein (5 copies)
-
|  
+
| packages with pIII
|-
|-
| VII  
| VII  
| 33
| 33
| phage head protein (5 copies)
| phage head protein (5 copies)
-
|  
+
| packages with p9
|-
|-
| VIII  
| VIII  
| 50
| 50
| phage coat protein (2700 copies)
| phage coat protein (2700 copies)
-
|  
+
| seg seq cleaved, matured form at aa 23 = clones to N-term, "Petrenko" site [[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&dopt=AbstractPlus&list_uids=8888146&query_hl=2&itool=pubmed_docsum]] 
|-
|-
| IX  
| IX  
| 32
| 32
| phage head protein (5 copies)
| phage head protein (5 copies)
-
|  
+
| stop codon overlaps start of gVIII
|-
|-
| X  
| X  
| 111
| 111
| DNA replication
| DNA replication
-
|  
+
| internal reinitiation inside gII
|-
|-
| XI  
| XI  
| 108
| 108
| assembly
| assembly
-
|  
+
| translation reinitiation inside gI
|}
|}
===Vectors for M13 protein expression===
===Vectors for M13 protein expression===
*M13KE [[http://www.westburg.nl/pages/TechnicalResources/Vectors.htm]]
*M13KE [[http://www.westburg.nl/pages/TechnicalResources/Vectors.htm]]
*pUC118 [[http://seq.yeastgenome.org/vectordb/vector_descrip/PUC118.html]] or 119 [[http://seq.yeastgenome.org/vectordb/vector_descrip/PUC119.html]]
*pUC118 [[http://seq.yeastgenome.org/vectordb/vector_descrip/PUC118.html]] or 119 [[http://seq.yeastgenome.org/vectordb/vector_descrip/PUC119.html]]
-
*M13K07 (helper phage)[[http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/sequences/m13ko7.txt]]
+
*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]]
==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===

Revision as of 10:28, 24 October 2006

Contents

Due Diligence

Life Cycle of the Phage

M13 phage proteins

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

Vectors for M13 protein expression

Makeover ideas

phage or phage coat rewrite

separate 8, 9, 3

two copies of 8

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