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Plasmids 101: Transformation, Transduction, Bacterial Conjugation, and Transfection

Posted by Alyssa Cecchetelli on Jun 25, 2019 8:54:52 AM

Horizontal gene transfer (HGT) is the movement of genetic material between organisms. It plays a key role in bacterial evolution and is the primary mechanism by which bacteria have gained antibiotic resistance and virulence. Scientists have studied how HGT occurs in nature and have learned how to introduce genetic materials into cells in the lab.

The introduction of foreign DNA or RNA into bacteria or eukaryotic cells is a common technique in molecular biology and scientific research. There are multiple ways foreign DNA can be introduced into cells including transformation, transduction, conjugation, and transfection. Transformation, transduction, and conjugation occur in nature as forms of HGT, but transfection is unique to the lab. Let’s take a look at these different methods of DNA insertion.

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Topics: Plasmids 101

Plasmids 101: Five Popular Model Organisms

Posted by Various Addgenies on Apr 11, 2019 8:39:27 AM

By Alyssa Cecchetelli and Lukas Morgan

Model organisms are vital tools used by researchers around the globe. These organisms share many genes with humans, are easily maintained in the lab, and have short generation times that make it easy to study the effects of genetic manipulations. In this blog post, we’ll cover five popular model organisms, but there are many more out there.

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Topics: Plasmids 101

Plasmids 101: Biotinylation

Posted by Alyssa Cecchetelli on Nov 15, 2018 8:50:12 AM

Biotin and its binding partner avidin are commonly used today in molecular biology for an array of different techniques and protocols. In this post we will discuss the natural role of biotin, biotinylation, the discovery of the biotin-avidin interaction and the uses of biotinylation in molecular biology!

Learn about in vivo biotinylation of bacterial fusion proteins

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Topics: Plasmids 101

Plasmids 101: Simplify Cloning with in vivo Assembly

Posted by Guest Blogger on Oct 18, 2018 8:37:05 AM

This post was contributed by Jake Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology.

Plasmid cloning is an essential part of any molecular biology project, yet very often, it is also a bottleneck in the experimental process. The majority of current cloning techniques involve the assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation. However, while not widely known, plasmid assembly can be achieved in vivo using a bacterial recombination pathway that is present even in common lab cloning strains.

This intrinsic bacterial recombination pathway, referred to as recA-independent recombination, joins together pieces of linear DNA through short homologous sequences at their termini, and likely functions as a bacterial DNA repair mechanism. The pathway is ubiquitous, with successful recombination reported in all laboratory E. coli strains tested so far.

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Topics: Plasmids 101, Plasmid Cloning

Plasmids 101: Codon usage bias

Posted by Tyler Ford on Sep 27, 2018 9:09:41 AM

A similar genetic code is used by most organisms on Earth, but different organisms have different preferences for the codons they use to encode specific amino acids. This is possible because there are 4 bases (A, T, C, and G) and 3 positions in each codon. There are therefore 64 possible codons but only 20 amino acids and 3 stop codons to encode leaving 41 codons unaccounted for. The result is redundancy; multiple codons encode single amino acids. Evolutionary constraints have molded which codons are used preferentially in which organisms - organisms have codon usage bias.

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Topics: Plasmids 101

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