Wyniki wyszukiwania

Wyszukiwano:
w słowach kluczowych: metabolic engineering

Sortuj według:

Ogranicz wyniki do:

Artykuł dostępny w postaci pełnego tekstu - kliknij by otworzyć plik

Transgenic callus culture establishment, a tool for metabolic engineering of Rhodiola rosea L.

100%

Mirmazloum I., Forgacs I., Zok A., Pedryc A., Gyorgy Z.

Acta Scientiarum Polonorum. Hortorum Cultus

2014

tom 13

nr 4

Agrobacterium tumefaciens EHA101 (pTd33) strain carrying uidA (GUS) reporter gene was used in model experiments on roseroot callus transformation. The T-DNA of pTd33 binary vector plasmid harbors nptII gene conferring resistance to kanamycin, and a uidA reporter gene, encodes the ß-glucuronidase enzyme. Roseroot seeds were sterilized and germinated on half strength MS media of which 70% germinated without any pretreatment. Calli were obtained from leaf segments of the in vitro grown seedlings. Calli was grown on solid MS medium supplemented with 1 mg l⁻¹ NAA and 0.5 mg l⁻¹ BAP. Different types of calli were obtained of which the green and compact type was chosen for transformation experiments. After co-cultivation with agrobacteria, calli were transferred to the same medium supplemented with 20 mg l⁻¹ kanamycin, 200 mg l⁻¹ carbenicillin and 300 mg l⁻¹ claforan with antioxidants (Polyclar and DTE) for selection. GUS test using a titron buffer was applied for monitoring the transformation of the calli. DNAs of 20 individual samples was extracted and subjected for PCR analysis proved the stable transformation in all of the taken samples by amplifying the nptII gene fragment. The method introduced here can be a tool for inserting and over-expressing the genes encoding for hypothesized enzymes to be involved in the biosynthesis of pharmaceutically important bioactive molecules of roseroot and therefore facilitating the applications for callus culture of roseroot in different bioreactor systems for pharmaceutical productions.

Alterations in protein secretion caused by metabolic engineering of glycosylation pathways in fungi

84%

Kruszewska J. S., Perlinska-Lenart U., Gorka-Niec W., Orlowski J., Zembek P., Palamarczyk G.

Acta Biochimica Polonica

2008

tom 55

nr 3

447-456

Due to its natural properties, Trichoderma reesei is commonly used in industry-scale production of secretory proteins. Since almost all secreted proteins are O-glycosylated, modulation of the activity of enzymes of the O-glycosylation pathway are likely to affect protein production and secretion or change the glycosylation pattern of the secreted proteins, altering their stability and biological activity. Understanding how the activation of different components of the O-glycosylation pathway influences the glycosylation pattern of proteins and their production and secretion could help in elucidating the mechanism of the regulation of these processes and should facilitate creation of engineered microorganisms producing high amounts of useful proteins. In this review we focus on data concerning Trichoderma, but also present some background information allowing comparison with other fungal species.

Ograniczanie wyników

1 Acta Biochimica Polonica

1 Acta Scientiarum Polonorum. Hortorum Cultus

1 Forgacs I.

1 Gorka-Niec W.

1 Gyorgy Z.

1 Kruszewska J. S.

1 Mirmazloum I.

1 Orlowski J.

1 Palamarczyk G.

1 Pedryc A.

1 Perlinska-Lenart U.

1 Zembek P.

1 Zok A.

1 2014

1 2008

Wyniki wyszukiwania

Transgenic callus culture establishment, a tool for metabolic engineering of Rhodiola rosea L.

Alterations in protein secretion caused by metabolic engineering of glycosylation pathways in fungi