Recombinant E. Coli
Recombinant E. Coli - Related Videos
07:35
Non-chromatographic Purification of Recombinant Elastin-like Polypeptides and their Fusions with Peptides and Proteins from Escherichia coli
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Cited by 48 •
2014
Elastin-like polypeptides are stimulus-responsive biopolymers with applications ranging from recombinant protein purification to drug delivery. This protocol describes the purification and characterization of elastin-like polypeptides and their peptide or protein fusions from Escherichia coli using their lower critical solution temperature phase transition behavior as a simple alternative to chromatography.
08:56
Recombineering and Gene Targeting
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2023
One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that allow them to be joined together. However, this technique has certain restrictions that limit its applicability for large or complex DNA construct generation. A newer technique that addresses some of these shortcomings is recombineering, which modifies DNA using homologous recombination (HR), the exchange between different DNA molecules based...
09:43
Purification and Refolding to Amyloid Fibrils of (His)6-tagged Recombinant Shadoo Protein Expressed as Inclusion Bodies in E. coli
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Cited by 3 •
2015
A two-step chromatographic method is described for the purification of recombinant Shadoo protein expressed as inclusion bodies in Escherichia coli, as well as a protocol to fibrillate purified Shadoo into amyloid structures.
05:19
Optimized Production and Analysis of Recombinant Protein-Filled Vesicles from E. coli
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Cited by 9 •
2023
The present protocol describes a detailed method for the bacterial production of recombinant proteins, including typically insoluble or disulfide-bond containing proteins, packaged inside extracellular membrane-bound vesicles. This has the potential to be applied to versatile areas of scientific research, including applied biotechnology and medicine.
03:40
Measuring E. coli Bacterial Load in Drosophila melanogaster following E. coli Infection
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2025
In this video, we demonstrate a technique for measuring the E. coli bacterial load after injecting E. coli into the thorax of Drosophila melanogaster.
FAQs
The plasmid serves as the DNA vehicle that carries the target gene into the bacterial cell. Once introduced, it can be maintained and replicated, allowing researchers to preserve the inserted sequence within a cell population. This makes plasmids central to gene cloning and supports later studies that require production of the encoded protein.
After transformation, selection identifies cells that maintain the recombinant plasmid. This step focuses further work on bacterial cells carrying the intended added DNA rather than on the entire transformed population. Maintaining those selected cells supports continued cloning and, when appropriate regulatory sequences are present, subsequent protein-expression experiments.
Regulatory sequences provide the signals needed for the inserted gene to be expressed in bacterial cells. When these sequences are suitable for the intended system, the engineered cells can produce the encoded protein rather than merely carry the DNA. This distinction allows the platform to support both gene cloning and protein expression.
By placing a target gene in a bacterial genetic context, researchers can maintain and replicate the sequence, clone it, and, when suitable regulatory sequences are present, produce its encoded protein. These capabilities provide a practical system for studying the gene and its product within molecular biology and biotechnology research.
Researchers first assemble the target gene into a plasmid, then introduce that recombinant plasmid into bacterial cells through transformation. They next select cells that maintain and replicate the added DNA. If the construct includes suitable regulatory sequences, the selected cells can also be used to produce the encoded protein.
These cells support several linked outcomes: cloning a target gene, maintaining recombinant DNA, expressing an encoded protein, and investigating gene function. Their use also extends to producing research reagents and medically relevant proteins. Consequently, the system connects basic molecular biology studies with practical biotechnology production.