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- W2392575807 abstract "Objective:To investigate the effects of the two expression cassettes of one vector on the level of the expression in different cell lines,to determine the optimal mutual position and transcriptional directions of light and heavy chain cassettes to the expression.Methods:Two complete antibody expression vectors including pIRESdhfrA and pIRESdhfrB,have been constructed.They differ from each other by their mutual position and transcriptional directions of light and heavy chain cassettes.The expression levels of the two vectors in CHO cells were compared via transient expression.Two derived vectors pIRESdhfrA-sv40ori and pIRESdhfrB-sv40ori were constructed and transfected to COS-7 cells.Their transient expression levels in COS-7 were evaluated by sandwich ELISA.To be able to compare the expression levels of the pIRESdhfrA and pIRESdhfrB in engineered CHO cells,two gene-targeting vectors,pIRESdhfrA-FRT and pIRESdhfrB-FRT were constructed.These gene-targeting vectors were then transfected to the engineered CHO cell line,and their expression levels were compared.Results:Transient expression in CHO cell line shows that the expression level of pIRESdhfrB was as 2.18 fold higher than the expression level of pIRESdhfrA.In comparison to the expression level of pIRESdhfrA-sv40ori,the expression level of the pIRESdhfrB-sv40ori was 2.3 fold higher in COS-7.Furthermore,the ratio of expression levels of the two engineered CHO cell line which were targeted with pIRESdhfrA-FRT and pIRESdhfrB-FRT was 11.6 to 1.Conclusion:The mutual position and transcriptional directions of the two expression cassettes in the vector pIRESdhfr constructed,head-to-head is better than head-to-tail for the expression of recombinant protein." @default.
- W2392575807 created "2016-06-24" @default.
- W2392575807 creator A5011787061 @default.
- W2392575807 date "2007-01-01" @default.
- W2392575807 modified "2023-09-28" @default.
- W2392575807 title "Effects on the Expression of Two Cassettes with Different Mutual Position and Transcriptional Directions" @default.
- W2392575807 hasPublicationYear "2007" @default.
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