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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/38446

Title: A transformation system for the nonuniversal CUG(Ser) codon usage species Candida rugosa.
Authors: Shye-Jye Tang
Kuang-Hui Sun
Guang-Huan Sun
Terng-Yuan Chang
Wan-Lin Wu
Guan-Chiun Lee
Contributors: 國立臺灣海洋大學:生命科學暨生物科技學系
Keywords: Transformation
Candida rugosa
Bleomycin-resistant determinant (ble)
Codon usage
Dominant selection
Zeocin
Date: 2003
Issue Date: 2016-09-01T08:35:06Z
Publisher: The Journal of Microbiological Methods
Abstract: Abstract: Since Candida rugosa utilizes a nonuniversal serine codon CUG rather than leucine, no vectors have been constructed to transform this organism. Moreover, it is difficult to design a new transformation system because no selection markers and promoters are available. In this study, Zeocin (400 μg/ml) was demonstrated to inhibit the growth of C. rugosa. The dominant selectable marker bleomycin-resistant determinant (ble) gene containing five CUG codons in an open-reading frame of 375 bp was synthesized by replacing its CUG codons into leucine codons (zeo-n). This marker conferred resistance to Zeocin. GAL1 promoter, transcription elongation factor 1 (TEF1) promoter from Saccharomyces cerevisiae and LIP3 promoter from C. rugosa were then used to drive zeo-n and to examine the function of promoter in C. rugosa. The resulting vectors enabled selection of Zeocin-resistant clones after transformation by LiCl method and electroporation. These results demonstrate that transformation into C. rugosa is feasible under the operation of GAL1, TEF1, and LIP3 promoters. The development of the transformation system for C. rugosa is essential to the genetic analysis of gene regulation and biochemical features of this fungal species and the expression of recombinant proteins in C. rugosa.
Relation: 52(2)
URI: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/38446
Appears in Collections:[生命科學系] 期刊論文

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