Abstract
An effective methodology is described for the direct expression of PCR-generated DNA fragments in cell-free transcription/translation systems without cloning DNA into plasmids. The methodology is accomplished here for the synthesis of the active antibacterial peptide cecropin using the synthetic coding DNA sequence. The proposed approach can be generally used for the cell-free synthesis of polypeptides and proteins that are unstable in living cells, or proteins that are strongly cytotoxic. Moreover, PCR-generated copies and/or multi-copies of genes and their fragments from genomic libraries (or genomic DNA) can be expressed directly in a transcription/translation system and then tested, without the sub cloning of DNA.
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© 2002 Springer-Verlag Berlin Heidelbreg
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Gudkov, A.T., Martemyanov, K.A. (2002). Direct Expression of PCR Products in Cell-Free Translation Systems. In: Spirin, A.S. (eds) Cell-Free Translation Systems. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59379-6_5
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DOI: https://doi.org/10.1007/978-3-642-59379-6_5
Publisher Name: Springer, Berlin, Heidelberg
Print ISBN: 978-3-642-63956-2
Online ISBN: 978-3-642-59379-6
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