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pGEM-T easy vector for expression


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6 replies to this topic

#1 penguin_th

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Posted 24 March 2005 - 06:47 PM

pUC19 is used for sequencing veactor and some time is used to expression vector (a little expression). pGEM-T easy veactor also has used for sequencing but can use for expressed the gene same with pUC19 or not. thank for answers.

#2 george@CASE

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Posted 28 March 2005 - 08:52 AM

what's the question?

#3 penguin_th

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Posted 28 March 2005 - 08:57 PM

Can usep GEM-T easy veactor for expressed the gene same with pUC19 or not?

#4 methylnick

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Posted 28 March 2005 - 10:13 PM

Can usep GEM-T easy veactor for expressed the gene same with pUC19 or not?

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I think your question is whether a gene ligated into pUC19 and can be expressed, be also done using pGEMT easy. If so......

pGEMTeasy has both T7 and Sp6 promoters so I am sure pUC19 also has these so I think the answer to your question........is yes

Nick

#5 penguin_th

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Posted 29 March 2005 - 12:57 AM

Thank you so much, Mr Nick..

#6 KevinK

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Posted 29 March 2005 - 09:37 AM

The pGEM-T Easy vector as it is supplied will actually NOT make a good expression vector. The reason is that this vector does not contain a ribosomal binding site and will therefore not translate well. If you clone an insert in to the vector and engineer a ribosome binding site, then verify that the gene is in the proper orientation to be transcribed by T7, the plasmid could be used for expression with a T7 expressing bacterial strain [i.e. BL21 (DE3)]. Unless all of these conditions are met though, the expression will be very poor. This would be the same for pUC series vectors as well by the way. I hope this helps. If you have any further questions, feel free to e-mail me at techserv@promega.com.

Regards,
Kevin

Technical Services Scientist
Promega Corporation
Promega Corporation
Madison, WI

#7 methylnick

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Posted 29 March 2005 - 09:37 PM

that's handy to know kevin, I use pGEMT easy for sequencing mainly.

Nick




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