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Codon Optimization | Expression Plasmids and Transient Transfections The Mayinga and Makona GP proteins contain amino acids 1-647 followed by a His6-tag. For the Mayinga and Sudan (Gulu) GP protein without the mucin-like domain, amino acids 320 until 476 were deleted and for the Makona protein without the mucin-like domain, amino acids 314 until 472 were deleted. DNA encoding the glycoproteins (GPs) were synthesized and codon-optimized for expression in human cells at GenScript (Piscataway, NJ 08854). | Get A Quote |
Ebola virus causes severe hemorrhagic fever, often leading to death in humans. The trimeric fusion glycoprotein (GP) is the sole target for neutralizing antibodies and is the major focus of vaccine development. Soluble GP ectodomains are unstable and mostly monomeric when not fused to a heterologous trimerization domain. Here, we report structure-based designs of Ebola and Marburg GP trimers based on a stabilizing mutation in the hinge loop in refolding region 1 and substitution of a partially buried charge at the interface of the GP1 and GP2 subunits. The combined substitutions (T577P and K588F) substantially increased trimer expression for Ebola GP proteins. We determined the crystal structure of stabil... More