3 replies to this topic

[moorele]

Hi,

I am looking for a vector within which I can put both my gene of interest and EGFP but under different promoters. My PI is pretty insistent that they're not under the same promoter eg gene of interest driven by CMV and EGFP driven by ubiquitin promoter. Does such a vector exist? I read something about bidirectional vectors where one promoter can drive the expression of 2 genes in different directions but I cant seem to find them - are they commercially available?

The vector will ultimately be used for transient transfection of cancer cell lines. Any help would be greatly appreciated.

Edited by moorele, 07 July 2009 - 04:33 AM.

[Dr Teeth]

Some of your message is unclear.  Do you want your 2 genes to be under different promoter control?  You mentioned bidirectional expression, which would be under the control of the same promoter.  I think what you want then is any vector with an internal ribosomal entry site (IRES) for bicistronic expression.  These are not bidirectional, but allow two products to be translated from a single mRNA.  Most investigators use these constructs when you want to ensure that if you see GFP, then your cell is also expressing your gene of interest.  Invitrogen makes many of these, as I am sure other companies do as well.  See: http://www.clontech....s...847&tabno=2.  It would be cheaper though to get a similar vector from Addgene.com

Edited by Dr Teeth, 07 July 2009 - 05:36 AM.

[moorele]

I think ideally they would like to have the 2 genes under different promoter control but an IRES vector may be an option. I will have a look at this! Thanks a mill

Dr Teeth, on Jul 7 2009, 02:33 PM, said:

Some of your message is unclear.  Do you want your 2 genes to be under different promoter control?  You mentioned bidirectional expression, which would be under the control of the same promoter.  I think what you want then is any vector with an internal ribosomal entry site (IRES) for bicistronic expression.  These are not bidirectional, but allow two products to be translated from a single mRNA.  Most investigators use these constructs when you want to ensure that if you see GFP, then your cell is also expressing your gene of interest.  Invitrogen makes many of these, as I am sure other companies do as well.  See: http://www.clontech....s...847&tabno=2.  It would be cheaper though to get a similar vector from Addgene.com

Edited by moorele, 07 July 2009 - 05:53 AM.

[moorele]

Think I may have found what I'm looking for - bidirectional expression systems from Clontech:
http://www.clontech....s...&prodorder=[Price]%20DESC#product_table

I'll let you know how i get one with these! Thanks for all the help.