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Expression of Cloned Genes in Using the Methanol-Inducible Promoter

Cold Spring Harb Protoc. 2021-01; 
Clara L Kielkopf, William Bauer, Ina L Urbatsch
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Gene Synthesis … for the small-scale expression test, the preferred method for yeast cell lysis is mechanical shearing, usually using glass beads … through de novo gene synthesis, although costly, is expedient and is offered by many providers (GeneArt, Life Technologies; GenScript, Entelechon; etc … Get A Quote

摘要

is a methylotrophic yeast capable of metabolizing methanol as its sole carbon source. Growth in methanol-containing medium results in dramatic induction of genes in the alcohol oxidation pathway including alcohol oxidase (AOX), formaldehyde dehydrogenase (FLD), and dihydroxyacetone synthase (DHAS). These proteins may comprise up to 30% of the biomass. Investigators have exploited these methanol-dependent genes to generate tightly regulated expression vectors. Most vectors use the strong and tightly regulated promoter to drive heterologous protein expression. Obtaining integrated transformants requires more DNA than transformations into , where the gene is expressed from episomal plasmids; however, transforma... More

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