TRANSFER DNA CINCHONA LEDGERIANA KEDALAM SEL SACCHAROMYCES CEREVISIAE

• Main Author: , Subandi
• Format: Theses
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/4472
• Institution: Institut Teknologi Bandung

The possibility for in vitro production of Cinchona alkaloid by the transformation of total Cinchona set DNA into Saccharomyces cerevisiae was tested. In this preliminary study the effect of DNA transfer into the yeast were observed by the... change in the morphology of the yeast transformOt cells i.e the shape and color, followed by the analyses of the secondary metabolites/ alkaloids produced when tryptophane was added as an inducer. <br /> This study can be devided into 3 parts first, the isolation of Cinchona DNA from Cinchona seedlings by the CTAB method; second the introduction of the DNA into the yeast cell by using the spheroplast method; and third the detection/ separation of transformed cells phenotype microscopically and the test for alkaloids produced using the TLC and HPLC tech nique. <br /> The DNA isolates were regarded as pure enough, based on the ratios of absorbans at 260 and 280 as 1.695. The rendement. of DNA isolate per seedling is 1,27 µg. <br /> The yeast transformant caused a difference in metabolite/ alkaloid composition in as well as out side the cells as compared to the control. This difference is manipested in terms of reduction of the peak for tryptophane in the medium and an increase in the cells as compared to the control, while the peaks for cinchona alkaloids increases in cell as well as in the medium. <br /> The transformation of DNA into the yeast does not change the phenotype as well as the appearence of the yeast cells.