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lactic acid bacteria - cloning

I m lost somewhere :(

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#1 rmbio

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Posted 18 November 2014 - 02:20 AM

Hi,

I wish to use/create food-grade expression system for lactic acid bacteria for expressing some genes of my interest into them. There are several such expression systems already described in the published papers. Although I have lot of experience in cloning and recombinant expression of genes, probably because of lot of reading I am totally confused, have lost the track and stuck in between.  My doubts are as following

 

1) Where can I get the plasmids from? Can I directly request the authors who have constructed them? If they give it to me and using them if I construct my own plasmids with some really useful genes having some commercial apllication and if I patent such new plasmids/recombinant strains, would there be any copyright/IP- related issues with the authors who shared the original plasmid with me?

 

2) Would it be a good idea to start de novo: to isolate naturally occuring plasmid in the species/strain that I am interested in and to generated a cloning verctor out of it for my purpose of food-grade expression?

 

3) If I just want some replication genes from these plasmids, can I simply amplify them from these plasmids so as to not have any copyrigth issue?

 

4) Is it posssible to contruct a new plasmid vector by amplifing the replication genes and Ori from one vector, selection gene from the other and add MCS by gene synthesis?

 

(The reason why I want to have my own expression system is (1) only few expression systems avaialble commercially (2) they are costly (3) If I develop a new expression system, that itself would be a good piece of work (4) I can patent such vector and put n-number of genes of my interest into it probably having nnumerous different applications)

 

Any other related suggestion welcome!!

 

Thanks


Edited by rmbio, 18 November 2014 - 02:48 AM.


#2 phage434

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Posted 18 November 2014 - 05:04 AM

Many authors will send plasmids if asked politely. Some distribute their plasmids from Addgene, which acts as a clearinghouse for distribution. Often these come with an MTA (material transfer agreement) which spells out what you are allowed to do with them. If I were you I would not spend time worrying about patents and rights at your stage. Get the work done, then worry about it. It will be harder than you think. Isolating natural plasmids may work, but is hard. Instead, I'd recommend mining Genbank, where sequences of plasmids are already deposited in most cases. Then, if you want to use a portion of one, you can synthesize it (DNA2.0, Genscript, Gen9, Synthetic Genomics) rather than worrying about getting it from someone.



#3 rmbio

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Posted 18 November 2014 - 08:17 PM

Many authors will send plasmids if asked politely. Some distribute their plasmids from Addgene, which acts as a clearinghouse for distribution. Often these come with an MTA (material transfer agreement) which spells out what you are allowed to do with them. If I were you I would not spend time worrying about patents and rights at your stage. Get the work done, then worry about it. It will be harder than you think. Isolating natural plasmids may work, but is hard. Instead, I'd recommend mining Genbank, where sequences of plasmids are already deposited in most cases. Then, if you want to use a portion of one, you can synthesize it (DNA2.0, Genscript, Gen9, Synthetic Genomics) rather than worrying about getting it from someone.

Alright phage434, thank you for the suggestions :)






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