Is phage display vector commercially available?
Nathan O. Siemers
nathan at protos.bms.com
Thu Aug 24 09:48:37 EST 1995
>>> On 22 Aug 1995 23:00:22 -0700, zchai at COBRA.PATH.MONASH.EDU.AU (Zhonglin Chai) said:
>> In article <firstname.lastname@example.org>
>> zchai at COBRA.PATH.MONASH.EDU.AU (Zhonglin Chai) writes:
>>> Hello there,
>>> I Wonder if there is any phage display expression vector (for cDNA
>>> library construction) commercially available? I appreciate any
>>> form of help.
>> I believe Stratagene (La Jolla, CA) has a vector called SurfZAP.
>> This is a variant on the lambdaZAP system. The single strand
>> rescue phagemid that is incorporated into the lambda vector (a kind
>> of pBluescript) has an M13 gene III segment downstream of the
>> polylinker. You ligate your inserts into the lambda, package, and
>> infect bacteria. After making the lambda library, use a helper
>> phage to rescue the phagemid as a filamentous phagemid with the
>> displayed fusion protein.
>> Daniel Kim ps. I have never used this system.
Z> Unfortunately this SurfZAP has the leader protein gene (M13 gene
Z> III) located downstream from the insert gene, which makes it very
Z> difficult for cDNA libraries. Most of the cDNA inserts (oligo(dT)
Z> primed and directionally cloned) have, at least, an in-frame stop
Z> codon. Hence, the leader protein won't be produced (no fusion
Z> protein produced) and the expressed foreign protein won't be
Z> displayed on the phage surface.
Z> Do you know any phage display vectors that have a leader protein
Z> upstream the cloning site?
One solution to this I have seen in the literature is to fuse the cdna
to fos, then have the phage coat protein fused to jun. the resulting
proteins assemble together and display on phage. Sorry, I don't have
the ref but it is recent (~2 years old).
Nathan O. Siemers, Ph.D.
Bristol-Myers Squibb Pharmaceutical Research Institute
3005 First Avenue, Seattle, WA 98121
(206) 727-3741 siemers at bms.com
"You want fries with that?"
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