Schwanniomyces occidentalis (Debaryomyces occidentalis ) is able to grow rapidly with high cell mass on cheap starch as a carbon source, produce strong amylolytic enzymes extracellularly and secrete large proteins without hyper-glycosylation and measurable extracellular proteases. Schw. occidentalis thus has a high potential as, a useful alternative to Saccharomyces cerevisiae in the production of heterologous proteins. However, the molecular study of Schw. occidentalis has been very limited due to the insufficient transformation system and lack of gene expression information. / A new transformation system of Schw. occidentalis has been developed. This system was based on vector YEp13 ( LEU2) and a stable leu auxotrophic mutant, Schw. occidentalis DW88, obtained by treating the yeast with 1-methyl-3-nitro-1-nitrosoguanidine. The transformation efficiency of YEp13 by spheroplast-mediating method was 103 transformants/mug DNA. The 2-mum replicon is proposed to be responsible for YEp13 replication in Schw. occidentalis. The YEp13 stability in Schw. occidentalis was low, but it kept its structure in the yeast, suggesting that Schw. occidentalis DW88 does not modify foreign DNA. / After analysis of 14 cloned Schw. occidentalis genes and comparison of associated genes from both Schw. occidentalis and S. cerevisiae, 25 codons were arbitrarily chosen as putative preferred codons for Schw. occidentalis. They are similar to those of S. cerevisiae, except for TTA for leucine, and AAA for lysine. Codon Bias Index (CBI), a criterion to evaluate gene expression, is calculated from preferred codons. A computer program (PCBI) which reads a gene containing introns was developed to quickly calculate CBI. / Schw. occidentalis DWSS should be a good host to produce and secrete heterologous proteins and the putative preferred codons and program PCBI can facilitate molecular study of Schw. occidentalis. (Abstract shortened by UMI.)
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.35955 |
| Date | January 1999 |
| Creators | Wang, Tsung-Tsan, 1959- |
| Contributors | Lee, Byong H. (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Doctor of Philosophy (Department of Food Science and Agricultural Chemistry.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001657200, proquestno: NQ50279, Theses scanned by UMI/ProQuest. |
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