5 replies to this topic

[Maverix]

Does anyone know how to get a plasmid map of a particular plasmid called pMSP12-GFP.
Also, how can you tell if the plasmid has E.coli promoter. Is there database of plasmids and a way to modify the antibiotic resistance cassettes in them?
If it has promoter for Mycobacteria and not for E.coli, does that mean it will not grow in the cell?

Thanks for your help!

[fishdoc]

Maverix, on Jul 29 2009, 08:43 AM, said:

Does anyone know how to get a plasmid map of a particular plasmid called pMSP12-GFP.
Also, how can you tell if the plasmid has E.coli promoter. Is there database of plasmids and a way to modify the antibiotic resistance cassettes in them?
If it has promoter for Mycobacteria and not for E.coli, does that mean it will not grow in the cell?

Thanks for your help!

Do you mean a promoter or an origin of replication? Since most cloning is done in lab E. coli strains, I think there are good odds the plasmid replicates in E. coli. However, if you have a publication source for that plasmid, read about how it was constructed, and if E. coli was ever used as a carrier.

If you mean a promoter and not an origin of replication, then that can be tricky. You may still be able to look at the source to determine where a gene and/or promoter came from.

As for modifying resistance cassettes, it definitely can be done, but with the multitude of plasmids available, there's likely one elsewhere that will give you the same thing you want in the way you want it.

What is it about this plasmid that you need it for?

[cotchy]

I think you should try to contact Lalita Ramakrishnan University of Washington, Seattle she seems to be the person who developed this particular plasmid

[Maverix]

cotchy, on Jul 30 2009, 06:29 AM, said:

I think you should try to contact Lalita Ramakrishnan University of Washington, Seattle she seems to be the person who developed this particular plasmid

Hey, Thanks for the tip, but do you think its okay to just send an email requesting the plasmid map?

[Maverix]

fishdoc, on Jul 29 2009, 07:33 AM, said:

Maverix, on Jul 29 2009, 08:43 AM, said:

Does anyone know how to get a plasmid map of a particular plasmid called pMSP12-GFP.
Also, how can you tell if the plasmid has E.coli promoter. Is there database of plasmids and a way to modify the antibiotic resistance cassettes in them?
If it has promoter for Mycobacteria and not for E.coli, does that mean it will not grow in the cell?

Thanks for your help!

Do you mean a promoter or an origin of replication? Since most cloning is done in lab E. coli strains, I think there are good odds the plasmid replicates in E. coli. However, if you have a publication source for that plasmid, read about how it was constructed, and if E. coli was ever used as a carrier.

If you mean a promoter and not an origin of replication, then that can be tricky. You may still be able to look at the source to determine where a gene and/or promoter came from.

As for modifying resistance cassettes, it definitely can be done, but with the multitude of plasmids available, there's likely one elsewhere that will give you the same thing you want in the way you want it.

What is it about this plasmid that you need it for?

Hey there, yes I mean the Ori of replicatio. Let me read it up in her article and see how it was constructed. And I need the plasmid map for restriction digestion to check its presence.
Thanks for your help, N

[bob1]

Most plasmids are on genbank!