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제안 방법
- , 2005). In this study, we obtained stable transgenic strains for the production of recombinant TPS using a heterologous expression. Further studies including the development of a method for measuring the activity of recombinant proteins generated by transformants are needed.
대상 데이터
- pastoris can be genetically manipulated to produce various compounds that are only produced naturally in higher eukaryotes. In this study, five TPS expressing transformants were obtained. We could confirm that genes of transgenic strains were expressed at transcription level.
- The linearized DNA was directly introduced without carrier DNA and the transformed cells, obtained by using an electroporation device, were plated on YPD medium supplemented with zeocin (100 μg/ml). Out of the more than 20 obtained candidates, five healthy appearing candidate single cells were selected. The small number of transformants can be explained by the scale of the culture, the amount of plasmid used, transformation without carrier DNA, and dilution during the plating step.
- P. brumalis was obtained from the Korean Collection for Type Cultures (KCTC46459). Fungi were inoculated onto potato dextrose agar (BD Difco, Franklin Lakes, NJ, USA) and were incubated in the stationary phase at 28°C for 7 days.
이론/모형
- 6 shows, the molecular weight of the secreted TPS was about 45 kDa. This was consistent with the predicted molecular weight computed using Compute pI/Mw tool. The following factors may have contributed to the faint protein band: gene copy number, site and mode of chromosomal integration of the expression cassette, 5′ and 3′ untranslated regions of mRNA, A+T composition of cDNA, transcription and translation blocks, nature of the secretion signal, endogenous protease activity, host strain physiology, media and growth conditions, and fermentation parameters (Dong et al.
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