Re: Chlamy Digest, Vol 94, Issue 1, pChlamy_4 vector

[Hamed Naghoosi]

Dear Dr. Mouradov
we have used the same vector with the same problem. though the yeild of
nuclear expression of recombinant protein is very low in chlamy. we could
detect our peptide by ELISA method, and the recombinant peptide content was
about 2 pg/ml. I think you'd better to concentrate your product before
further analysis if you can not use ELISA.

best

Hamed Naghoosi
PhD Candidate in Microbiology,
Iranian Research Organization for Science and Technology (IROST).

On Sat, Oct 22, 2016 at 8:34 PM, <chlamy-...@oat.bio.indiana.edu> wrote:

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> 1. pChlamy_4 vector (Elizabeth Harris)
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> Message: 1
> Date: Sat, 22 Oct 2016 11:38:42 +0000
> From: Elizabeth Harris <chl...@duke.edu>
> Subject: [Chlamydomonas] pChlamy_4 vector
> To: "chl...@net.bio.net" <chl...@magpie.bio.indiana.edu>
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> Begin forwarded message:
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> From: Aidyn Mouradov <aidyn.m...@rmit.edu.au<mailto:
> aidyn.m...@rmit.edu.au>>
> Date: October 21, 2016 7:44:05 PM EDT
>
> Subject: pChlamy_4 vector
>
> Dear all
> We are using a pChlamy_4 vector (GeneArt® Chlamydomonas Protein Expression
> Kit) for expression of recombinant protein in chlamy cells. This vector
> contains a translational fusion between the coding region of zeocin gene
> (selectable marker) and the gene of interest with 2a peptide from foot and
> mouth disease in between (for post-translational separation of GOI). We are
> having a good number of transgenic lines grown on zeocin 10 confirmed with
> PCR and RT-PCR (to proof expression) but cannot get any proteins on
> Westerns. I am wondering if anyone had this type of problem with pChlamy_4
> vector or this is an effect of the cloned gene: a synthetic codon optimized
> version of the coat protein
> Many thanks
> Aidyn
>
> Associate Professor Aidyn Mouradov
> Plant and Algal Biotechnology
> Royal Melbourne Institute of Technology University
> RMIT Bundoora (West) campus, Bundoora VIC 3083, Australia
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