Molecular Cloning & Protein Expression
The aim of molecular cloning is to insert the gene-of-interest (GOI) into a plasmid vector, a circular piece of DNA that contains various elements to facilitate cloning, clone selection, and protein expression. Researchers often use DNA restriction enzymes and ligase to insert the GOI in-frame within the expression vector for subsequent protein expression. This vector is then inserted into a cell that will express the protein encoded by the GOI or protein expression is accomplished using cell-free protein synthesis systems.
Once the protein is expressed, the protein function can be studied as it affects the cell signaling, morphology, or other aspects within the cell. Alternatively, the protein can be expressed in large quantities, purified, and used in numerous downstream applications. For additional information, explore our comprehensive offering of reliable reagents and resources that are available across the cloning & expression workflow for insect, bacteria, and mammalian protein expression systems.
Products
RNA and DNA Modifying Enzymes
During molecular cloning, researchers frequently use restriction enzymes to cut DNA and other enzymes like RNA and DNA modifying enzymes to add, remove, or alter chemical groups on nucleic acids. To meet diverse research needs, we offer a comprehensive range of these essential enzymes including Alkaline Phosphatase, Protease, T7 RNA Polymerase, Pyrophosphatase, Ribonuclease A, and T4 DNA Ligase. Our reagents, combined with specially formulated buffers, help ensure high efficiency and reliability in DNA and RNA manipulation.
Novagen® pET System, Simplicon® Vectors, and UCOE® Protein Expression Systems
Our E. coli, insect, and mammalian expression vectors simplify the selection of positive transformants. Some highlighted products include:
- The Novagen® pET System is recognized as the gold standard for cloning and expressing recombinant proteins in bacterial systems.
- The Simplicon® In Vivo Protein Expression System offers a non-genome integrating, tunable, and synthetic self-replicating RNA-based expression system. Simplicon® expression generates immediate, high-sustained protein expression of multiple genes in transfected human cells without the risk of genome integration.
- The Ubiquitous Chromatin Opening Element UCOE® expression vector technology achieves grams of protein in less than 30 days in stably transfected mammalian cells. UCOE® expression technology prevents transgene silencing and gives more consistent, stable, and higher-level gene expression (up to 20-fold) than conventional vectors, regardless of the chromosomal integration site.
Bacterial and Yeast Transformation Reagents
Bacterial transformation involves inserting foreign DNA into bacteria using methods such as heat shock and electroporation. Our portfolio of competent cells is engineered for optimal plasmid and protein expression, even for challenging experimental conditions. Highlights include:
- NovaBlue® competent cells are optimized for efficiency and plasmid preservation, supporting superior library preparations and plasmid stability.
- Rosetta™, Origami™, and Overnight Express™ systems enhance proper protein folding and increase solubility or expression of cytotoxic proteins.
For yeast transformation, we provide straightforward protocols and scalable products that integrate seamlessly into protein expression workflows.
For mammalian cell research, check out our Transfection page for additional options.
Plasmid Vectors from BCCM/GeneCorner
Explore our expanded offering of plasmid vectors from BCCM/GeneCorner - a unique, ISO certified plasmid repository, hosted by the Department of Biomedical Molecular Biology of the Faculty of Sciences at Ghent University (UGent). This diverse collection includes a variety of cloning and recombinant protein expression plasmids in an array of target organisms, supporting modernized research in molecular and protein biology.
Each Product sale subject to the terms and conditions of Sigma-Aldrich’s Terms and Conditions for Sale to End Users Purchasing Third-Party Products and any statutory, regulatory or other restriction imposed by law.
- You may only use the Products in any lawful manner for research, teaching or quality control purposes. Any commercial use of the Products requires the prior written authorization of the Provider and may be subject to a written agreement. Such approval will not be unreasonably withheld. Commercial use means the use of the Products for the purpose of profit (such as the resale, leasing, exchange, license, or other transfer of the Products for profit purposes and any use of the Products to perform service activities for profit purposes).
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Related Resources
- Brochure: Molecular Essentials Guide
Learn how our comprehensive molecular tools and reagents provide precise, ready-to-use solutions for your workflows.
- Article: Cell-Free Protein Synthesis or Expression
Cell-free protein synthesis rapidly produces recombinant proteins without cells, utilizing translation machinery from different cell types for efficiency.
- Article: Competent Cell Selection & E. coli Markers Guide
Select competent cells by application, transformation efficiency, and genotype. We also present a list of E. coli markers for reference.
- Article: Simplicon™ in vivo Protein Expression System
Simplicon: A non-genome integrating, tunable and sustained protein expression system in human cells, based on a single, polycistronic, and self-replicating synthetic RNA.
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