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stupid question on cloning


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2 replies to this topic

#1 lna

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Posted 13 November 2009 - 05:05 AM

Hi everyone!
I assume that my question may sound weird.
But when you need to express some protein in eukaryotic cells (by transfecting them with the plasmid) - how do you clone the gene into your plasmid?
Do you take the WHOLE sequence of cDNA from gene bank to insert into the MCS of the vector or you can amplify just the part of it (making sure that it contains tha CODING sequence for the protein)?
;)

#2 elpollodiablo

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Posted 13 November 2009 - 05:29 AM

Hi everyone!
I assume that my question may sound weird.
But when you need to express some protein in eukaryotic cells (by transfecting them with the plasmid) - how do you clone the gene into your plasmid?
Do you take the WHOLE sequence of cDNA from gene bank to insert into the MCS of the vector or you can amplify just the part of it (making sure that it contains tha CODING sequence for the protein)?
;)


You just need the coding sequence to express the protein.

#3 lna

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Posted 13 November 2009 - 06:47 AM

Hi everyone!
I assume that my question may sound weird.
But when you need to express some protein in eukaryotic cells (by transfecting them with the plasmid) - how do you clone the gene into your plasmid?
Do you take the WHOLE sequence of cDNA from gene bank to insert into the MCS of the vector or you can amplify just the part of it (making sure that it contains tha CODING sequence for the protein)?
;)


You just need the coding sequence to express the protein.



oh, may be I'm not that hopeless..
thanks!




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