KR880007730A - Expression vector - Google Patents

Abstract

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Description

Expression vector

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Claims (18)

(a) an enhancer / promoter unit (which unit is functionally linked to (b) a fragment), (b) can encode a desired protein product, and contains signals for polyadenylation and effective transport to the cytoplasm DNA fragments capable of being transcribed into mRNA, followed by (c) promoter unit (C ₁ ) which is functionally linked to (C ₂ ) small fragments) and DNA small fragments (C which can be transcribed into mRNA of the selectable marker gene) ₂ ) a series of functional elements arranged in the order of a DNA fragment consisting of ₂ ), wherein the promoter unit (C ₁ ) lacks a functional enhancer but enhances the enhancer termination DNA between (a) and (C ₁ ) A recombinant DNA molecule that can be stimulated by the absence of a sequence and an enhancer present in (a). The method of claim 1, wherein the unit (C ₂ ) can be readily determined from an antibiotic resistant protein, an enzyme that inhibits the nutritional demand variability of the cell, or a group of enzymes that produce a changed color substrate from a colorless or different colored substrate. DNA molecule encoding a marker protein. The DNA molecule of claim 1, wherein the DNA fragment (C ₂ ) is from a prokaryote. The DNA molecule of claim 1, wherein the DNA fragment (C ₂ ) is from a eukaryotes. 4. The DNA molecule of claim 3, wherein the promoter / enhancer element (a) is present after the enhancer-end sequence. The DNA molecule of claim 5, wherein the enhancer-terminating sequence consists of DNA derived from a prokaryote. The DNA molecule of claim 4, wherein the promoter / enhancer element (a) is present after the enhancer-ending DNA sequence and the DNA fragment (c) is positioned before the enhancer-ending DNA sequence. 8. The DNA molecule of claim 7, wherein the enhancer-terminating sequence consists of DNA derived from a prokaryote. The DNA molecule of claim 1, wherein the marker gene (C ₂ ) encodes the enzyme dihydrofolate reductase. The method of claim 1, wherein the DNA sequence of unit (b) is serum protein, polypeptide hormones, hormonal receptors, lymphokines, interferons, enzymes, enzyme inhibitors, analgesic polypeptides, glycoproteins, immunoglobulins, immunoglobulins- A recombinant DNA molecule encoding a desired protein selected from a binding protein, an antigenic protein, a vaccine protein, a plasminogen activator or a precursor group of the target protein. The method according to claim 1, wherein the promoter / enhancer (a) is an actin gene, a metallothionein gene, other cellular genes, an SV40 early gene, a herpes virus gene, a human immediate early gene. , A recombinant DNA molecule containing a promoter and an enhancer selected from the group consisting of early genes immediately before the mouse cytomegalovirus strain and other viral genes. The method according to claim 1, wherein the promoter (C ₁ ) is a specific promoter or the SV40 early gene, the late adenovirus gene, the herpes thymidine kinase gene, the initial gene just before the human cytomegalovirus strain, the early gene immediately before the mouse cytomegalo virus strain. And a recombinant DNA molecule containing a promoter fragment selected from a gene selected from the group consisting of a cellular gene and a metallothionein gene. A host transformed with the DNA molecule of claim 1. The host of claim 13, wherein said host is selected from the group consisting of E. coli, other bacteria, yeasts, fungi, green plants, and animal cells. The host of claim 13, wherein said host is a Chinese hamster ovary cell line. The host of claim 13, wherein said host lacks the enzyme dihydro folate reductase. A method of producing a protein of interest, comprising culturing the transgenic host of claim 13 and isolating said protein. The method of claim 17 comprising culturing a mixed clone of the transformed host cell. ※ Note: The disclosure is based on the initial application.