Soft agar assays were conducted following transfection of cells with mammalian pCMV6-Entry expression vectors containing a G418 resistance cassette (Origene).
Whereas EIs are likely to be more effective in blocking infection of cells with low entry receptor expression levels, the other drugs, whose effectiveness is independent of the entry receptor expression level, are expected to block the infection of cells with high entry receptor expression levels, bringing about synergy.
Based on previous studies, 20, 26 we let S i depend on entry receptor expression following a Hill function,, where is the Hill coefficient and is the entry receptor expression level at which.
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It involves the formation of a cationic lipid DNA complex that interacts efficiently with the cell surface, leading to the entry and expression of the exogenous DNA.
The attachment of the peptide consisted of 12 amino acids on the C-terminus of adenovirus fiber protein significantly improved entry and expression of a β-galactosidase transgene into neuroglial cells such as astrocytes, and Schwann cells.
Two key steps are involved in the cationic lipid mediated gene transfer process: (1) interaction between the cationic liposomes and DNA leading to the formation of a complex and (2) interaction between the complex and the target cells leading to DNA entry and expression of the transgene.
All entry and expression recombinant clones are stored as DNA and as bacterial glycerol stocks (15% v/v glycerol) of single colony-selected E. coli transformants.
Experiments that measure the effectiveness of drugs as a function of entry receptor expression levels would provide tests of our proposed explanation.
The generated resources, including cDNA entry clones, expression constructs and transgenic plants, will facilitate further functional analysis of the members of this important gene family.
