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Optimization of in vitro translation using different human cell lines
Author(s) -
Dhodda Vinay K,
Jensen Penny,
Deshpande Priya,
Mcbride Aaron,
Kilmer Gregory,
Hommema Eric,
Choi Jae,
Webb Brian,
Vattem Krishna M.
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb50
Subject(s) - hela , translation (biology) , in vitro , cell culture , microbiology and biotechnology , cell , protein biosynthesis , biology , computational biology , chemistry , messenger rna , biochemistry , gene , genetics
Culturing of mammalian cells for the purpose of protein expression is a time consuming and expensive process. An effective alternative to cell culture is cell‐free protein expression (i.e., in vitro translation) using extracts prepared from mammalian cells. We have used several immortalized human cell lines, including HeLa and HepG2, to make translation competent extracts ideal for expressing proteins. We have also optimized the HeLa in vitro translation system for higher protein expression. To highlight the flexibility of our translation system, we have demonstrated expression of 5 different proteins in a single reaction. We have also shown the compatibility of our system with high throughput protein expression using a PCR‐based approach. These results indicate the utility of the HeLa in vitro translation system for expression of functional human proteins.