Chemistry: molecular biology and microbiology – Process of mutation – cell fusion – or genetic modification – Introduction of a polynucleotide molecule into or...
Reexamination Certificate
1999-12-07
2001-10-16
Wang, Andrew (Department: 1635)
Chemistry: molecular biology and microbiology
Process of mutation, cell fusion, or genetic modification
Introduction of a polynucleotide molecule into or...
C435S471000, C435S006120, C536S023100, C536S023200, C536S023700, C536S024100, C536S024300, C536S024320
Reexamination Certificate
active
06303381
ABSTRACT:
TECHNICAL FIELD
The present invention relates to a novel insertion sequence derived from a methane-assimilating bacterium. Because the insertion sequence may be transposed or inserted into various sites on chromosomes, it can be utilized as effective means for genetic analysis or means for incorporating a desired gene into a bacterial chromosome.
BACKGROUND ART
As for both of prokaryotes and eukaryotes, DNA sequences transposable from one site to another site within a genome or between different genomes have been known, and generally called transposable elements. Most of such transposable elements encode, within their own sequences, a site-specific recombination enzyme (transposase) required for the translocation from one site to another site. Among such transposable elements, those having the smallest and simplest structure are called insertion sequence (IS).
Insertion sequences among microbial transposable sequences exhibit several characteristics. They have a size of about 1 to 2 kb, and have inverted repeat sequences of about 8-20 bp at the both ends. When an insertion sequence is inserted into a microbial chromosome, duplication of a target sequence occurs on the end side of the insertion sequence (Mobile Genetic Elements, Academic Press, New York, p.159-221 (1983)). Among microbial insertion sequences, well studied are those derived from
Escherichia coli
[Mol. Gen. Genet., Vol. 122, 267-277 (1973)], those derived from Shigella [
J. Bacteriol.,
172, 4090-4099 (1993)], those derived from acetic acid bacteria [
Mol. Microbiol.,
9, 211-218 (1993)], those derived from mycoplasma [
Mol. Microbiol.,
7, 577-584 (1993)] and the like.
On the other hand, as for methane-assimilating bacteria, there have been known inventions of a method for continuous production of oxidation products utilizing such bacterial cells (Japanese Patent Laid-open No. 5-3279, Japanese Patent Laid-open No. 54-3583), environmental cleanup through degradation of environmental pollutants utilizing methane-assimilating bacteria (Japanese Patent Laid-open No. 6-245761, Japanese Patent Laid-open No. 8-24905), fermentation method for converting methane into materials for protein production (Japanese Patent Laid-open No. 50-40788) and the like. Thus, they are industrially important microorganisms. However, any insertion sequence derived from methane-assimilating bacteria has not been reported so far.
SUMMARY OF THE INVENTION
During the cloning of a gene coding for methane monooxygenase (it may be abbreviated as “sMMO gene” hereinafter) for functional analysis of the methane monooxygenase (it may be abbreviated as “sMMO” hereinafter), which is an enzyme derived from a methane-assimilating bacteria,
Methylococcus capsulatus,
the inventors of the present invention found that an insertion sequence was contained in that gene. The present invention has been accomplished based on this finding.
That is, the present invention provides an insertion sequence which has the nucleotide sequence shown in SEQ ID NO: 1 derived from chromosomal DNA of a Methylococcus bacterium. The present invention also provides an insertion sequence which has the inverted repeat sequences at the both ends, wherein each of the inverted repeat sequences consists of a sequence of the nucleotide numbers 5-19 of SEQ ID NO: 1.
REFERENCES:
T.M. Barta, et al., Antonie Van Leeuwenhoek Int. J. Gen. Mol. MicroBiol., vol. 64, No. 2, pp. 109-120, “Genetics of Methane and Methanol Oxidation in Gram-Negative Methylotroph Bacteria,” 1993.
A.C. Satinthorpe, Gene, vol. 91, pp. 27-34, “The Methane Monooxygenase Gene Cluster of Methylococcus Capsulatus (Bath),” 1990.
Gunji Yoshiya
Kawahara Yoshio
Sugimoto Shinichi
Yasueda Hisashi
Ajinomoto Co. Inc.
Oblon & Spivak, McClelland, Maier & Neustadt P.C.
Shibuya Mark L.
Wang Andrew
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