کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
24848 43541 2008 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
High-level expression of soluble viral structural protein in Escherichia coli
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
High-level expression of soluble viral structural protein in Escherichia coli
چکیده انگلیسی

Pharmaceutically relevant virus-like particles (VLPs) can potentially be manufactured cheaply and efficiently through in vitro assembly of viral structural protein in cell-free reactors, but a bottleneck for this processing route is the currently low-level expression of soluble viral protein in efficient cell factories such as Escherichia coli (E. coli). Here, we report expression levels of up to 180 mg L−1 that are achievable from low-cell-density E. coli cultures using a simple and low cost strategy. We investigated effects of host strain, plasmid, inducer concentration, pre-induction temperature and cell density at induction with design of experiment (DOE). The statistical approach successfully identified significant effects and their interactions, and provided insights into the role of codon-usage effects in expression of viral structural protein. In particular, our results support the notion that full codon optimization may be unnecessary to improve expression of viral genes rich in E. coli rare codons; using a strategically modified host cell could provide a simpler and cheaper alternative.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Biotechnology - Volume 134, Issues 1–2, 20 March 2008, Pages 64–71
نویسندگان
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