Skip to main content
. Author manuscript; available in PMC: 2017 May 1.
Published in final edited form as: Protein Expr Purif. 2015 Dec 29;121:17–21. doi: 10.1016/j.pep.2015.12.001

Figure 5.

Figure 5

Anti-GST Western analysis of wild-type and engineered cryptic site GSThRCC1HIS variant protein expressed in E. coli. (a) C-terminally HIS-tagged GSThRCC1 in crude extracts (lanes 1, 3) or metal affinity purified (lanes, 2, 4) complexes coexpressed in BL21(DE3)pLysS was detected using anti-GST antibodies. The wild-type GST sequence was used in lanes 1 & 2, while the GST sequence mutated to remove the cryptic Shine-Dalgarno mutation or START codon or both elements are shown in lanes 3 & 4 respectively. from the engineered cryptic translation initiation site was not detected (b) Amino acid and DNA sequences for the wild type and cryptic initiation site GST mutant.