User:Nkuldell:M13 redesign: Difference between revisions
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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 07:28, 24 October 2006
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 |
