Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Reexamination Certificate
2000-04-25
2004-03-02
Achutamurthy, Ponnathapu (Department: 1652)
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
C435S183000, C435S193000, C435S252300, C435S320100, C536S023200
Reexamination Certificate
active
06699689
ABSTRACT:
TECHNICAL FIELD AND BACKGROUND ART
The present invention relates to the recombinant DNA technology. The present invention also relates to the mass-production of antimicrobial materials from microorganisms and a DNA construct and vector system. Biologically active peptide (antimicrobial peptide hereinafter) has little chance to develop resistance since the antimicrobial peptides show activity by a mechanism that is totally different from that of conventional antibiotics which have a serious problem of developing resistance. Therefore, the antimicrobial peptides have a high industrial applicability in the fields of pharmaceutics and the food industry.
The main obstacle in the industrial use of the antimicrobial peptide, however, is the difficulty in economical mass-production of the antimicrobial peptides. For instance, the production of the antimicrobial peptides by chemical synthesis is not economical. Also, there have been attempts to produce antimicrobial peptides by genetic engineering using microorganisms, in this case, however, the expression levels of the antimicrobial peptides are very low.
U.S. Pat. No. 5,206,154 provides a DNA construct which comprises a polypeptide gene which is capable of suppressing the bactericidal effect of cecropin, and a cecropin gene fused to the polypeptide gene. An example of such polypeptide disclosed in the patent is the araB gene.
U.S. Pat. No. 5,593,866 provides a method for a microbial production of a cationic antimicrobial peptide, wherein the cationic peptides is expressed as a fusion to an anionic peptide to avoid degradation by a bacterial protease.
DISCLOSURE OF THE INVENTION
The present invention provides a DNA construct to mass-produce a antimicrobial peptides. The present invention also provides a DNA construct that can produce and recover antimicrobial peptides effectively from microorganisms.
Also, the present invention provides gene multimers that can increase the efficiency of expression, separation and purification of desired peptides and the construction method of such construct.
Further, the present invention provides an expression vector to mass-produce antimicrobial peptides from microorganisms.
Further, the present invention provides a method to mass-produce antimicrobial peptides form microorganisms.
REFERENCES:
patent: 5206154 (1993-04-01), Lai et al.
patent: 5593866 (1997-01-01), Hancock et al.
J. Yun Tso et al., “Nucleotide Sequence ofEscherichia colipurF and Deduced Amino Acid Sequence of Glutamine Phosphoribosylpyrophosphate”, The Journal of Biological Chemistry, vol. 257 No. 7 pp. 3525-3531 (1982).
Christopher A. Makaroff, “Cloning of theBacillus subtilisGlutamine Phosphoribosylpyrophosphate Amidotransferase Gene inEscherichia coli”, The Journal of Biological Chemistry, vol. 258 No. 17 pp. 10586-10593 (1983).
J.H. Lee, et al., “Acidic Peptide-Mediated Expression of the Antimicrobial Peptide Buforin II as Tandem Repeats inEscherichia Coli”, Protein Expr. Purif. Feb. 1998 (abstract).
L. Zhang et al., “Determinants of Recombinant Production of Antimicrobial Cationic Peptides and Creation of Peptide Variants in Bacteria”, Biochem. Biophys. Res. Commun., Jun. 29, 1998, (abstract).
M. Okomoto et al., “Enhanced Expression of an Antimicrobial Peptide Sarcotoxin A by GUS Fusion in Transgenic Tobacco Plants”, Plant Cell Physiol. Jan. 1998 (abstract).
C. Haught et al., “Recombinant Production and Purification of Novel Antisense Antimicrobial Peptide inEscherichia Coli”, Biotechno. Bioeng. Jan. 1998 (abstract).
Hong Seung Suh
Kang Min Hyung
Kim Jeong Hyun
Lee Hyun Soo
Lee Jae Hyun
Achutamurthy Ponnathapu
Darby & Darby
Fronda Christian L.
Samyang Genex Corporation
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