Extended gene expression by medium exchange and repeated transient transfection for recombinant protein production enhancement

2015 ◽  
Vol 112 (5) ◽  
pp. 934-946 ◽  
Author(s):  
Laura Cervera ◽  
Sonia Gutiérrez-Granados ◽  
Nicholas Simon Berrow ◽  
Maria Mercedes Segura ◽  
Francesc Gòdia
Keyword(s):  
Gene Expression ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Transient Transfection ◽  
Medium Exchange

Construction of a binary transgenic gene expression system for recombinant protein production in the middle silk gland of the silkworm Bombyx mori

2009 ◽  
Vol 19 (3) ◽  
pp. 473-487 ◽  
Author(s):  
Ken-ichiro Tatematsu ◽  
Isao Kobayashi ◽  
Keiro Uchino ◽  
Hideki Sezutsu ◽  
Tetsuya Iizuka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Recombinant Protein ◽  
Bombyx Mori ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Expression System ◽  
Silk Gland ◽  
Gene Expression System ◽  
Silkworm Bombyx Mori

Transient gene expression: Recombinant protein production with suspension-adapted HEK293-EBNA cells

2001 ◽  
Vol 75 (2) ◽  
pp. 197-203 ◽  
Author(s):  
Petra Meissner ◽  
Horst Pick ◽  
Alexandra Kulangara ◽  
Philippe Chatellard ◽  
Kirstin Friedrich ◽  
...  
Keyword(s):  
Gene Expression ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Transient Gene Expression

scholarly journals Optogenetic lac operon to control chemical and protein production in Escherichia coli with light

2019 ◽  
Author(s):  
Makoto A. Lalwani ◽  
Samantha S. Ip ◽  
Cesar Carrasco-Lopez ◽  
Evan M. Zhao ◽  
Hinako Kawabe ◽  
...  
Keyword(s):  
Gene Expression ◽  
Metabolic Engineering ◽  
Recombinant Protein ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Lac Operon ◽  
Iptg Induction ◽  
Spatial Control ◽  
Chemical Inducers ◽  
Cell Functions

AbstractControl of the lac operon with IPTG has been used for decades to regulate gene expression in E. coli for countless applications, including metabolic engineering and recombinant protein production. However, optogenetics offers unique capabilities such as easy tunability, reversibility, dynamic induction strength, and spatial control that are difficult to obtain with chemical inducers. We developed an optogenetic lac operon in a series of circuits we call OptoLAC. With these circuits, we control gene expression from various IPTG-inducible promoters using only blue light. Applying them to metabolic engineering improves mevalonate and isobutanol production by 24% and 27% respectively, compared to IPTG induction, in light-controlled fermentations scalable to at least 2L bioreactors. Furthermore, OptoLAC circuits enable light control of recombinant protein production, reaching yields comparable to IPTG induction, but with enhanced tunability of expression and spatial control. OptoLAC circuits are potentially useful to confer light controls over other cell functions originally engineered to be IPTG-inducible.

Transient Transfection Factors for High-Level Recombinant Protein Production in Suspension Cultured Mammalian Cells

2008 ◽  
Vol 39 (2) ◽  
pp. 141-153 ◽  
Author(s):  
Chaoting Liu ◽  
Brian Dalby ◽  
Weixing Chen ◽  
Jennifer M. Kilzer ◽  
Henry C. Chiou
Keyword(s):  
Recombinant Protein ◽  
Mammalian Cells ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Transient Transfection ◽  
High Level

scholarly journals Development of a constitutive and an auto-inducible high-yield expression system for recombinant protein production in the microalga Nannochloropsis oceanica

2020 ◽  
Vol 104 (20) ◽  
pp. 8747-8760
Author(s):  
Imke de Grahl ◽  
Sweta Suman Rout ◽  
Jodi Maple-Grødem ◽  
Sigrun Reumann
Keyword(s):  
Gene Expression ◽  
Nitrate Reductase ◽  
Recombinant Protein ◽  
Reporter Gene ◽  
Recombinant Protein Production ◽  
Protein Production ◽  
Reporter Gene Expression ◽  
Total Soluble Protein ◽  
High Yield ◽  
Nannochloropsis Oceanica

Abstract Photoautotrophic microalgae offer a great potential as novel hosts for efficient recombinant protein production. Nannochloropsis oceanica produces an extraordinarily high content of polyunsaturated fatty acids, and its robust growth characteristics, published genome sequence and efficient nuclear transformation make N. oceanica a promising candidate for biotechnological applications. To establish a robust and flexible system for recombinant protein production, we cloned six endogenous, potentially constitutive or inducible promoters from N. oceanica strain CCMP1779 and investigated their strength using monomeric Venus as reporter gene. Microscopic pre-screening of individual transformants revealed that the promoters of elongation factor (EF), tubulin (TUB) and nitrate reductase (NR) enabled high reporter gene expression. Comparative quantitative analyses of transformant populations by flow cytometry and qRT-PCR demonstrated the highest Venus expression from the EF promoter and the NR promoter if extended by an N-terminal 14-amino acid leader sequence. The kinetics of reporter gene expression were analysed during photobioreactor cultivation, achieving Venus yields of 0.3% (for EF) and 4.9% (for NR::LS) of total soluble protein. Since inducible expression systems enable the production of toxic proteins, we developed an auto-induction medium for the NR promoter transformants. By switching the N source from ammonium to nitrate in the presence of low ammonium concentrations, the starting point of Venus induction could be fine-tuned and shifted towards exponential growth phase while maintaining high recombinant protein yields. Taken together, we demonstrate that a model recombinant protein can be produced robustly and at very high levels in N. oceanica not only under constitutive but also under auto-inducible cultivation conditions. Key points • Nannochloropsis oceanica might serve as host for recombinant protein production. • Comparative promoter strength analyses were conducted for twelve different constructs. • Robust high-yield recombinant protein production was achieved under constitutive conditions. • The nitrate reductase promoter enabled protein production under auto-induction conditions.

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