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Protein Expression
Click for a larger view. (http://www.niehs.nih.gov/research/atniehs/core/pe/images/slide1a.gif) Using PCR to make the entry clone results in the greatest number of options when deciding how to express the protein. The design of the PCR product will dictate if the desired gene can be expressed with n or c terminal tags, or no tags at all. It is also the only way to introduce proteolytic cut sites in most of the expression vectors. The entry clone can also be made by traditional sub cloning and TOPO® cloning methods. 
Click for a larger view. (http://www.niehs.nih.gov/research/atniehs/core/pe/images/slide1b.gif) |
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