Impaired dominant selectable marker sequence and intronic insertion strategies for enhancement of expression of gene product and expression vector systems comprising same
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United States of America Patent
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Dec 12, 2000
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app pub date -
Mar 30, 1999
filing date -
Nov 13, 1992
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Abstract
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 'x' is a nucleotide selected from the group consisting of adenine (A), quanine (G), cytosine (C) or thymine (T)/uracil (U); 'Py' 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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