Impaired dominant selectable marker sequence and intronic insert

Details Impaired dominant selectable marker sequence and intronic insert Impaired dominant selectable marker sequence and intronic insert

1993-11-03

1997-07-15

Fleisher, Mindy

Chemistry: molecular biology and microbiology

Vector, per se

4352523, 435358, 435325, 435357, 435352, 435354, 435369, 435372, 435364, 435365, C12N 1563, C12N 121, C12N 506, C12N 510

Patent

active

056482671

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: 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.

REFERENCES:
Yenofsky et al., 1990, PNAS, 87:3435-3439.
Abrams et al, 1989, J. Biol. Chem., 264(24):14016-14021.
Kozak, 1989, J. Cell Biol. 108:229-240.
Kozak, 1986, PNAS., 83:2850-2854.
Kozak, 1987, Mol. Cell. Biol., 7(10):3438-3445.
Kozak, 1984, Mol. Cell. Biol., 9(11):5073-5014.
Niwa, 1991, Gene, 108:193-200.
Sambrook et al., 1989, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Press: Cold Spring Harbor N.Y; selected unnumbered pages.
Hartman et al, PNAS, 85:8047-8051, 1988.

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