In an era of designer mice and complex multi-component DNA constructs, it is difficult to imagine that there was a time (not so long ago) when biologists lacked the ability to manipulate DNA sequences. The dawn of the modern molecular biology era was brought about by a series of influential innovations known as the Molecular Biology Revolution. Here are a few of them:
In the early 1950s, two groups described “restriction factors” in bacteria that prevented bacteriophage infection1,2, but it wasn’t until 15 years later that the first restriction enzymes were isolated by Arber and Linn, and independently by Smith and Wilcox3,4. Since then, numerous restriction enzymes have been isolated and become commercially available.
In the early 1960s, the work of Jean Weigle, Matthew Meselson, and Grete Kellenberger demonstrated that DNA molecules could reassemble by ligation5,6. In 1967, enzymes with this ligase activity were isolated by several groups7-11. For the first time, DNA fragments could be assembled in a test tube in specific arrangements.
The power of restriction enzymes and ligases was quickly harnessed by Paul Berg, who assembled the first truly recombinant DNA molecules using DNA from E. coli, bacteriophage and Simian virus 4012. At nearly the same time, Cohen et al. performed the first complete demonstration of the power of modern molecular biology: They used restriction digestion, ligation, and transformation to transfer an engineered, functional DNA molecule into a bacterial strain13.
In the early 1980s, two key advances were made that revolutionized molecular biology even further. First, Marvin Caruthers developed phosphoramidite DNA synthesis, which made automated oligonucleotide synthesis practical14,15. Second, Kary Mullis published polymerase chain reaction (PCR)16. Using oligo synthesis and PCR together, researchers suddenly had the ability to selectively amplify, and therefor clone, virtually any target DNA sequence. The use of many oligos with complimentary overlaps, coupled with ligation and PCR, also allowed large DNA fragments with de novo sequence to be created from scratch.
Most of the innovations leading to the Molecular Biology Revolution are now considered common knowledge among biologists. Custom DNA constructs have become ubiquitous tools in numerous types of experiments, and the assembly of a recombinant plasmid is no longer considered a significant scientific achievement. In fact, many labs have begun using external resources, such as core facilities, plasmid repositories, and commercial cloning services for their cloning needs, abandoning molecular cloning in their lab altogether.
VectorBuilder is the latest addition to the Molecular Biology Revolution. It is a highly innovative web tool that allows you to design complex custom DNA vectors with just a few mouse clicks. You can then purchase the physical vector for as little as $100, and get it mailed to your lab in week or two. You can choose from a wide range of vector systems, including regular plasmids, lentiviral vectors, shRNA vectors, CRISPR/Cas9 vectors, and many more.
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