Impaired dominant selectable marker sequence and intronic insertion strategies for enhancement of expression of gene product and expression vector systems comprising same
Disclosed herein are fully impaired consensus Kozak sequences which are most typically used with dominant selectable markers of transcriptional cassettes which are a part of an expression vector. These vectors are most typically utilized in the expression of proteins in mammalian expression systems. As defined, disclosed and claimed herein, a \"fully impaired consensus Kozak\" comprises the following sequence: ##STR1## where: Nat nucleotides 2,3,8 and 9 is a nucleotide selected from the group consisting of adenme (A), quanine (G), cytosine (C) or thymine (T)/uracil (U); Nat nucleotides 1 and 7 is a pyrimidine nucleotide, ie C or T/U; \"ATG\" is a codon encoding for the amino acid methionine, the so-called \"start\" codon; and -3 and +1 are directional reference points vis-a-vis ATG, ie -3 is meant to indicate three nucleotides upstream of ATG and +1 is meant to indicate one nucleotide downstream of ATG. Dominant selectable markers further comprising artificial intronic insertion regions are further disclosed.
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