Heterologous production of five Hepatitis C virus-derived antigens in three Saccharomyces cerevisiae host strains.
In this study, the production of recombinant Hepatitis C virus (HCV) derived proteins from transformed Saccharomyces cerevisiae yeast cells is reported. Three different yeast strains (GRF18U, BY4743-4A and CENPK 113-5D) have been transformed for the intracellular expression of five antigens of different dimensions (from 32.8 to 85.2 kDa), all derived from the non-structural (NS) region of different HCV viruses' genotypes and posed under the control of a glycolytic promoter. The putative trans-membrane domains contained in four antigens seem responsible of their accumulation as protein aggregates. Good productions of the smaller and of the bigger antigens (50 and 30 mgl(-1), respectively) have been observed in simple flask batch cultures. Productions are strongly dependent from the genetic background of the yeast host and from the cellular localization of the antigen, while they appear independent from the growth rate of the transformed hosts. For every recombinant antigen tested, the highest production levels were achieved with the CENPK 113-5D-host strain, while the GRF18U strain shows symptoms of a heavily stressed phenotype.
Parolin
Carola
C
Università degli Studi di Milano - Bicocca, Dipartimento di Biotecnologie e Bioscienze, p.za della Scienza 2, 20126 Milano, Italy.
Corso
Andrea Dal
AD
Alberghina
Lilia
L
Porro
Danilo
D
Branduardi
Paola
P
D018936:Hepatitis C Antigens / Q000096:biosynthesis* / Q000235:genetics; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics; D013045:Species Specificity; D014162:Transfection / Q000379:methods
D018936:Hepatitis C Antigens; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins