Research Article

Influence of Codon Bias on Heterologous Production of Human Papillomavirus Type 16 Major Structural Protein L1 in Yeast

Figure 2

Analysis of HPV-16 L1 expression in yeast by SDS-PAGE (a) and Western blot with mouse polyclonal antibody against HPV-16 L1 protein (b). The same samples were run on each gel. In lanes: (1) crude lysate of yeast expressing ORF L1-Pv; (2) crude lysate of yeast expressing ORF L1-Ec; (3) crude lysate of yeast expressing ORF L1-Pl; (4) crude lysate of yeast expressing ORF L1-Hm; (5) crude lysate of yeast expressing ORF L1-Sc; (6) negative control sample from crude lysate of S. cerevisiae cells transformed with the empty vector pFX7; and M: prestained protein weight marker (Thermo Scientific Fermentas). Long arrow points to the band with HPV16 L1 protein. The protein band (~50 kDa) pointed with short arrow was used to determine the ratio of this protein in the lines for evaluation of loaded yeast lysates quantitative differences using the ImageQuant TL 1D gel analysis software (GE Healthcare). The ratio of ~50 kDa protein band in lines was 1.77, 1.00, 1.69, 1.74, 1.30, and 4.44 accordingly. The data from one representative experiment are shown. The expression level of L1 proteins in 10 randomly picked transformants in every group was alike.
979218.fig.002a
(a)
979218.fig.002b
(b)