Cell-free expression is a convenient
method for rapidly producing protein. Coupled transcription-translation
systems provide the advantage of using plasmid DNA as the temple for
cell-free expression with no need to first transcript the DNA. The
researcher not only has flexibility in which template to use, but also in
which system to use as there are now mammalian, plant, bacterial and insect
cells systems. Protein detection is a requirement for most applications.
Traditionally, one would add radioactive [35S]-methionine to their cell-free
expression reactions. With increased regulations on radioactivity,
researchers are now using other detection methods such as Fluorotech,
Transcend, and Western blotting. Researchers are using cell-free expressed
proteins in many applications including protein:protein pulldowns,
protein:nucleuic acid shift assays, and enzymatic assays. With new systems
that can produce large amounts of protein, researchers are expanding this
list of applications.
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