pYFP vector - what does what (Oct/25/2006 )

hi guys
most of the time when we use a vector we don't look it in details.
so I ask a lot of question a vector call peYFPN1:

-it's start by a pCMVIE then the MCS and the YFP.
I guess the PCMV is the promoter for YFP.correct?

-then come after that the SV40PolyA: I guess is for the processing of the RNA YFP. correct?

-then a f1ori: ?

-then a P"?

-then a SV40ori/pSV40e:?

-then the Kan/Neo

-then HsVTK Poly A: I guess for the Kan/neo RNA:

-then a pUC ori:?

any help?

thanks

jm

-ulujm-

This is a Clontech pEYFP-N1 vector - its description you can find here http://orders.clontech.com/clontech/techin.../pEYFP-N1.shtml
Il

-ili_-

QUOTE (ili_ @ Oct 25 2006, 12:19 PM)

This is a Clontech pEYFP-N1 vector - its description you can find here http://orders.clontech.com/clontech/techin.../pEYFP-N1.shtml
Il

I know but they don't go in details.
for exemple why 2 ori: pucori and f1 ori.

-ulujm-

1) pUC plasmid replication origin
2) f1 single-strand DNA origin (Packages the noncoding strand of EYFP)

-ili_-

1) pUC plasmid replication origin
2) f1 single-strand DNA origin (Packages the noncoding strand of EYFP)
[/quote]

yes so can you explaine what it means

thanks

-ulujm-

Replication origins - regions where DNA synthesis is initiated.
Three Common Features of Replication Origins:
1) replication origins are unique DNA segments that contain multiple short repeated sequences;
2) these short repeat units are recognized by multimeric origin-binding proteins - these proteins play a key role in assembling DNA polymerases and other replication enzymes at the sites where replication begins;
3) origin regions usually contain an AT-rich stretch. This property facilitates unwinding of duplex DNA because less energy is required to melt A·T base pairs than G·C base pairs.

-ili_-

f1 single-strand DNA origin - origin of replication of bacteriophage f1 --> http://www.pubmedcentral.nih.gov/articlere...gi?artid=431910

-ili_-

Hi thanks
so the ppuc orig is usefull to replicate the plasmid in the bacteria I presume.
When we grow the bacteria the plasmid inside will replicate.
I don't see the use of the f1 ori replication of bacteriophage

jm

-ulujm-

The f1 ori is very useful!! It allows the formation of phage-like particles (of course after the infection with helper phage) containing single-stranded DNA what is necessary of Kunkel mutagenesis method.

-ili_-