pChlamy_4 vector

[Elizabeth Harris]

Begin forwarded message:

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
Editorial board member of journals:
Journal of Sustainable Bioenergy Systems
Source Journal of Bioremediation
NanoPhotoBioSciences



T: + (61) 3-9925-7144| E: aidyn.m...@rmit.edu.au<mailto:aidyn.m...@rmit.edu.au>
http://www.rmit.edu.au/contact/staff-contacts/academic-staff/m/mouradov-associate-professor-aidyn<https://urldefense.proofpoint.com/v2/url?u=http-3A__www.rmit.edu.au_contact_staff-2Dcontacts_academic-2Dstaff_m_mouradov-2Dassociate-2Dprofessor-2Daidyn&d=CwMFaQ&c=imBPVzF25OnBgGmVOlcsiEgHoG1i6YHLR0Sj_gZ4adc&r=tztUD9iumFyQ37MEXMtAjg&m=HsKOadRc-HCKAQMgt-3ec3NpTe2v4a7W8Sgl2fL30to&s=pxAvy-qkEAF4CLvLoR1VCMIQCPhdf3rQmUx8DksJWJA&e=>



Even if you're on the right track, you'll get run over if you just sit there."
--Will Rogers