Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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Expression of Cloned Genes in E. coli Using IPTG-Inducible Promoters

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Many E. coli expression vectors use regulatory elements derived from the lac operon, which is unsurprising given that the lac operon represents a paradigm for prokaryotic gene regulation (for review, see Reznikoff 1992). Because the lac promoter itself is relatively weak, strong hybrid promoters have been developed for production of foreign proteins in E. coli (for review, see Baneyx 1999). As described in the introduction to this chapter, promoters for the expression of foreign proteins in E. coli usually depend on either the host cell RNA polymerase or on introduction of a foreign polymerase derived from the bacteriophage T7. Both types of vector are induced by addition of the nonhydrolyzable lactose analog isopropyl--D-thiogalactopyranoside (IPTG). The protocol given here is intended for use with these and other IPTG-inducible vectors. Other systems are described in the Discussion section.


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(Limited time special offer.) Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook
Molecular Cloning Fourth Edition, A Laboratory Manual, by Michael R. Green and Joseph Sambrook

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