GATEWAY-CLONING - Experience with GATEWAY and exchange of clones (Jun/29/2005 )

The Gateway system from Invitrogen is used by many groups with rather different results. It is difficult to find people to exchange Gateway vectors and experience.
Anybody interested in sharing vectors and experience is highly wellcome - please post your message here.
My lab has to offer: Entry-vectors (ENTR1A and DONR221), the adenoviral system (CMV-promotor and promotorless), the TRex-system for inducible expression and Block-It vectors.
We are looking for: lentiviral vector

Exchange of vectors and experience is officially encouraged by Invitrogen (see their homepage).

Best
Stefan

-stefan-

Hehehe...

I was recently at a conference where I got to hear both sides of the story about Gateway cloning, my sympathies are with you!

Dr Washo

-dr_washo-

QUOTE (stefan @ Jun 29 2005, 11:36 AM)

-stefan-

Dr. Washo,

could you please be more specific?

Thanks
Stefan

-stefan-

I am encountering some problems with gateway vectors. I am using PENTRA-1 and destination vector 42 for cloning flue matrix protein cDNA. I discovered on N-terminal sequencing of the protein made, that it starts about 9 amino acids ahead of the`designated start codon. Translation starts at TTG codon in the vector sequence. I am wondering if it is normal to encounter these problems with gateway vectors or something wrong with my experiments.

regards

Seema

-skhan-

QUOTE (stefan @ Jun 29 2005, 03:36 AM)

I know it has been a while since you posted this -- I am new to this forum. However, I am interested in sharing vectors. I have pLenti6/V5-DEST already, and would like to use this backbone for shRNA. However I would need both pDONR221 vector and Block-It vector for this. What do you say??
Thanks!
Chandreyee Das
Dana-Farber Harvard Cancer Center

-cdas-