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Sh ble and Cre adapted for functional genomics and metabolic engineering of Pichia stipitis

Paper ID Volume ID Publish Year Pages File Format Full-Text
18389 42721 2006 7 PDF Available
Title
Sh ble and Cre adapted for functional genomics and metabolic engineering of Pichia stipitis
Abstract

Pichia stipitis is widely studied for its capacity to ferment d-xylose to ethanol. Strain improvement has been facilitated by recent completion of the P. stipitis genome. P. stipitis uses CUG to code for serine rather than leucine, as is the case for the universal genetic code thereby limiting the availability of heterologous drug resistance markers for transformation. Development of a modified selectable marker for resistance to bleomycin (Sh ble) and efficient excision of the marker after integration (loxP/Cre) should facilitate functional genomics and metabolic engineering in this yeast. The Sh ble marker did not code for an active protein in P. stipitis until four CUG codons were mutagenized to TTG, which is properly translated as leucine in yeasts that use the alternative yeast nuclear genetic code. The 18 CTG codons in Cre were mutagenized in a similar manner and the system was used to delete XYL2. The resulting xyl2Δ mutant did not use xylose as a carbon source.

Keywords
Transformation; Genetic engineering; Yeast; Expression; Mutagenesis; Sh ble; Cre; Alternative yeast nuclear genetic code; CUG
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Sh ble and Cre adapted for functional genomics and metabolic engineering of Pichia stipitis
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Publisher
Database: Elsevier - ScienceDirect
Journal: Enzyme and Microbial Technology - Volume 38, Issue 6, 1 April 2006, Pages 741–747
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us