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- W2393088529 abstract "To develop an robust strategy for the tandem repeat co-expression of multiple genes in one plasmid,the point mutations were introduced into commercial expression vector pET-22b by the overlap PCR.After the mutation,the restriction enzyme site XbaⅠ was moved to the upstream of T7 promoter and a SpeⅠ site was introduced to the downstream of T7 terminator.The whole expression cassette including the T7 promoter,ribosome binding sites,multiple cloning site,T7 terminator were covered by the XbaⅠ-SpeⅠrestriction fragment.Restriction digestion and sequencing indicate that the mutated expression vector pET-m22b was constructed successfully.The four genes of different sources were choose for the coepxression by the pET-m22b.The SDS-PAGE shown that the four genes were highly co-expressed and without any affection on each other.The results indicated an efficient co-expression vector suitable for the tandem combination expression of multiple genes in single plasmid was constructed,which will make the co-expression of many genes more easy and will also greatly facilitate the expression and purification of multi-subunit protein complex,the rapid heterogenesis re-construction and optimization of biochemical pathways etc." @default.
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- W2393088529 date "2011-01-25" @default.
- W2393088529 modified "2023-09-24" @default.
- W2393088529 title "Construction of a Vector Suitable for the Tandem Coexpression of Multiple Genes by a Single Plasmid" @default.
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