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were first optimized for high level protein expression in E. coli. They were then synthesized using DNA oligo based, PCR gene assembling method. Poly-arginine tag plus a linker sequence, i.e. ESGGGGSPGRRRRRRRRRRR, was added to each protein C-terminal. The final DNA fragment was flanked with Nde I and Xho I sites, and inserted into pET41a expression vector(More)
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