Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1992-09-30
1995-10-24
Draper, Garnette D.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
4352401, 43525233, 4353201, 536 231, C12N 1509, C12N 1517
Patent
active
054609542
ABSTRACT:
The specification describes a process for producing human proinsulin in Escherichia coli (E. coli) using gene manipulation technology. The process can provide for human proinsulin in high yields by a novel expression vector having strong regulatory elements of an insulin gene and a stable recombinant gene product. The expression vector of the present invention is characterized in that: 1) it has an 11 amino acid leader peptide containing six threonines in order to ensure an intracellular stability of proinsulin fusion protein, 2) it contains two copies of a DNA expression cassette each comprising a strong lambda P.sub.R promoter, a lac ribosome binding site, a proinsulin gene with a 17 amino acid leader peptide sequence containing a DNA sequence encoding (Thr).sub.6, and a strong fd phage transcription terminator (combination of phage fd terminator and translation stop codon), etc. successively ligated, 3) it has an ampicillin resistance gene, 4) it can be very stably retained within a cultured cell, and 5) there are a number of these expression vectors in E. coli by which the expression can be significantly increased. Human insulin is prepared from the proinsulin fusion protein by in vitro conversion.
REFERENCES:
patent: 5017478 (1991-05-01), Cashion et al.
patent: 5102789 (1992-04-01), Siegal et al.
Kang et al., 1991, Biotechnology Letters, 13, 755-760.
Goeddel et al., "Expression in Escherichia coli of Chemically Synthesized Genes for Human Insulin" Proc. Natl. Acad. Sci. USA 76(1):106-110 (1979).
Chance et al., "Chemical, Physical, and Biological Properties of Biosynthetic Human Insulin" Diabetes Care 4(2):147-154 (1981).
Williams et al., "Cytoplasmic Inclusion Bodies in Escherichia coli Producing Biosynthetic Human Insulin Proteins" Science 215:687-689 (1982).
Frank et al., "The Production of Human Proinsulin and its Transformation to Human Insulin and C-Peptide" In: Peptide (eds. Rich and Gross, Dierce Chemical Company, Rockford, Ill.) pp. 729-738 (1981).
Guo et al., "Synthesis of Human Insulin Gene" Gene 29:251-254 (1984).
Sung et al., "Short Synthetic Oligodeoxyribonucleotide Leader Sequences Enhance Acculation of Human Proinsulin Synthesized in Escherichia coli" Proc. Natl. Acad. Sci. USA 83:561-565 (1986).
Laemmli, U. K., "Cleavage of Structure Proteins during the Assembly of the Head of Bacteriophage T4" Nature 227:680-685 (1970).
Kang Yup
Kim Choong S.
Lee Hyune S.
Lee Hyune W.
Lee Jae H.
Cheil Foods & Chemicals Inc.
Draper Garnette D.
Teng Sally
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