SummaryTransformation techniques ge

Summary
Transformation techniques generally require development before genetic and
molecular studies of industrial yeast strains can commence. Candida glycerinogenes
WL2002-5 has been used for industrial-scale glycerol production but has proven
difficult to transform for molecular studies following previously published
procedures. In the present study, phleomycin was used as drug-resistance marker
based on perturbing plasma membranes and the lower minimum inhibitory
concentration for C. glycerinogenes. We developed an efficient method for
transformation of C. glycerinogenes, and parameters involved in transformation
efficiency were optimized. Pretreatment of yeast cells with either dithiothreitol or
lithium acetate enhanced the frequency of transformation markedly by electroporation.
Significantly higher transformation efficiency was obtained when the
electroporated cells were pretreated with phleomycin before plating. With this
method, a maximal transformation frequency of 394750 transformants/mg plasmid
DNA was obtained. The transformation method will foster genetic manipulation of
this industrial strain.

Hasil (Bahasa Indonesia) 1:

RingkasanTransformasi teknik umumnya memerlukan pembangunan sebelum genetik danmolekul studi strain ragi industri dapat dimulai. Candida glycerinogenesWL2002-5 telah digunakan untuk produksi skala industri gliserol tapi telah terbuktisulit untuk mengubah molekul studi berikut diterbitkan sebelumnyaprosedur. Dalam penelitian ini, phleomycin digunakan sebagai penanda resistensi obatBerdasarkan perturbing membran plasma dan minimum yang lebih rendah penghambatankonsentrasi untuk C. glycerinogenes. Kami mengembangkan metode yang efisien untuktransformasi C. glycerinogenes, dan parameter terlibat dalam transformasiefisiensi yang dioptimalkan. Pretreatment ragi sel dengan baik dithiothreitol ataulithium asetat ditingkatkan frekuensi transformasi nyata oleh elektroporasi.Secara signifikan lebih tinggi efisiensi transformasi diperoleh ketikasel-sel electroporated yang pretreated dengan phleomycin sebelum plating. Dengan inimetode, frekuensi maksimal transformasi 394750 plasmid transformants mgDNA diperoleh. Metode transformasi akan memupuk manipulasi genetikstrain ini industri.