Recombinant Expression
Recombinant Expression - Related Videos
09:46
Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris
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Cited by 59 •
2010
The protocol describes protein expression using the methylotrophic yeast Pichia pastoris. The preparation of electrocompetent yeast cells, transformation of the vector with the gene of interest into P. pastoris and yeast DNA purification are also performed. Western blot analysis and protein purification build the last steps in this protein expression protocol.
12:18
Expression of Functional Recombinant Hemagglutinin and Neuraminidase Proteins from the Novel H7N9 Influenza Virus Using the Baculovirus Expression System
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Cited by 182 •
2013
Here we describe a way to express correctly folded and functional influenza virus surface antigens derived from the novel Chinese H7N9 virus in insect cells. The technique can be adapted to express ectodomains of any viral or cellular surface proteins.
07:50
Efficient Agroinfiltration of Plants for High-level Transient Expression of Recombinant Proteins
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Cited by 200 •
2013
Plants offer a novel system for the production of pharmaceutical proteins on a commercial scale that is more scalable, cost-efficient and safe than current expression paradigms. In this study, we report a simple and convenient, yet scalable approach to introduce target-gene containing Agrobacterium tumefaciens into plants for protein transient expression.
08:09
High-throughput Titration of Luciferase-expressing Recombinant Viruses
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Cited by 14 •
2014
This article presents a high-throughput luciferase expression-based method of titrating various RNA and DNA viruses using automated and manual liquid handlers.
03:44
Generation of GFP-Tagged Recombinant Respiratory Syncytial Virus Using a T7 Expression System
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2026
Source: Bouillier, C. et. al., Amplification, and Titration of Recombinant Respiratory Syncytial Viruses. J. Vis. Exp. (2019)This video demonstrates the generation of a green fluorescent protein (GFP)-expressing recombinant respiratory syncytial virus using a T7-based reverse genetics system. The co-transfection of plasmids encoding viral proteins and a GFP-tagged genome leads to protein expression and initiates virus rescue.
FAQs
Promoters and other regulatory elements determine how the inserted coding sequence is handled by the host cell. In an expression vector, these elements provide the control framework that allows host machinery to transcribe the selected sequence and translate its product. Their inclusion links recombinant DNA design to the amount and usability of protein available for later analysis or purification.
Host choice changes the cellular setting in which transcription and translation occur. Bacteria, yeast, and mammalian cells can all serve as production platforms, but each supplies its own host machinery and culture environment. In bioengineering, selecting among these systems helps align the recombinant construct with the intended goal, such as protein production, structural study, or functional analysis.
The expression vector organizes the coding sequence with regulatory elements needed for expression. This arrangement creates a controllable interface between recombinant DNA and the host cell. Consequently, vector design is central to converting a selected sequence into a recoverable gene product rather than leaving the sequence without an expression context suitable for transcription and translation.
A typical workflow begins by selecting the coding sequence, placing it in an expression vector with regulatory elements, and transferring the construct into a chosen host cell. Researchers then maintain the host under selected culture conditions so the gene product is made, followed by harvesting and purification. This sequence connects molecular design, cellular production, and downstream recovery.
Culture conditions matter because recombinant production depends on the host environment as well as the DNA construct. Researchers can control the conditions under which bacteria, yeast, or mammalian cells maintain expression, then evaluate the resulting product for recovery or study. Managing both the host system and culture setting supports scalable production and more consistent outcomes.
After expression, harvesting and purification produce a preparation of the gene product suitable for further use. The recovered material can support structural studies, functional analysis, or development of therapeutics, industrial enzymes, vaccines, and diagnostic reagents. Downstream recovery therefore connects cellular production with the practical form required for research, biotechnology, or product development.
In bioengineering, recombinant expression connects engineered DNA to practical biomolecule development. The strategy can provide material for structural studies and functional analysis while also supporting scalable manufacture of therapeutics, industrial enzymes, vaccines, and diagnostic reagents. Its value comes from combining genetic construct design with controllable biological production and recovery of the resulting gene product.