Impaired dominant selectable marker sequence and intronic insert

Chemistry: molecular biology and microbiology – Vector – per se

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536 231, 536 241, C12N 1585

Patent

active

057337799

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.

REFERENCES:
Yenofsky, 1990, Natl. Acad. Sci., 87: 3435-3439.
Abrams, 1989, J. Biol. Chem., 264: 14016-14021.
Kozak, 1989, J. Biol. Chem., 108: 229-241.
Kozak, 1986, Proc. Natl. Acad. Sci., 83: 2850-2854.
Kozak, 1987, Mol. Cell Biol., 7(10): 3438-3445.
Kozak, 1984, Mol. Cell Biol., 9(11): 5073-5084.
Niwa, 1991, Gene, 108: 193-200.
Sambrook et al., 1989, Molecular Cloning: A Laboratory Manual, Colo. Springs/Harbor Press, Colo. Spring Harbor NY.
Hartman et al., 1988, Proc. Natl. Acad.Sci., 85: 8047-8051.

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