Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1996-05-24
1998-12-15
Ketter, James
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 711, 4351723, 4351735, 4353201, 43525422, 536 231, 536 241, 536 242, C12P 2102
Patent
active
058495240
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to reproducible transformation systems of Candida utilis, more particularly to the transformation of the yeast Candida utilis with recombinant DNA, and to the expression of heterologous genes in novel transformants obtained thereby. The present invention also relates to novel DNA sequences which may be used as selectable marker genes for transformation, and to novel DNA sequences which may be used as promoters or terminators for expressing heterologous genes. In addition, the present invention relates to methods for the efficient integration of heterologous genes into yeast chromosomes.
Furthermore, the present invention relates to DNA fragments having the properties of autonomous replication as well as enhancing transformation efficiency in Candida utilis, to methods for integrating DNA fragments having no selectable marker gene into chromosome, and to methods for obtaining DNA sequences having promoter activity.
2. Background Art
The development of gene manipulation technology has made it possible to produce useful proteins in a large amount with microorganisms. Prokaryotes such as Escherichia coli or Bacillus subtilis can be used easily as a host therefor, inter alia, E. coli being employed most frequently as a host. However, proteins produced in E. coli are often led to insoluble forms and cannot be glycosylated upon secretion, so that these proteins are not satisfactory for a variety of requisitions. In addition, pyrogenic toxic factors produced by E. coli must be removed, when proteins thus produced are intended to be used as medicaments.
As compared with the use of these prokaryotic systems, it is interesting to use eukaryotes such as yeast as a host for producing useful proteins. First of all, yeasts of the genus Saccharomyces or the yeast Candida utilis have been known to be of a high safety, since Saccharomyces has been long used for the production of fermentation products such as alcoholic products and the yeast Candida utilis has been used for the production of feeds. Moreover, yeast can be generally cultured at a cell density higher than bacteria as well as in a continuous mode. Yeast secretes proteins into a medium, and the protein secreted is modified by glycosylation. The production of proteins with yeast is thus of worth when such modification is important for biological activity of the protein.
Yeasts classified in the genus Saccharomyces have been investigated most extensively, and genetic information of them have been accumulated. The yeasts have been investigated as a host for producing a variety of substances. Further, transformation technology for some of yeasts in addition to Saccharomyces such as those in Pichia, Hansenula, Kluyveromyces, Candida genera has been developed, and these yeasts are now examined as a host for producing useful substances. Among them, the yeasts assigned to the genus Candida, inter alia have properties advantageous for practical use which are not found in the yeasts of the Saccharomyces genus such as a wide anabolic spectrum of the carbon source. The yeast of the genus Candida is thus expected to be used for the production of useful substances with recombinant DNA technology.
Among the yeasts of the genus Candida, Candida utilis has an excellent anabolic ability to pentoses such as xylose. In addition, Candida utilis, different from the yeast of the genus Saccharomyces, does not produce ethanol in culture under aerobic conditions and thus the growth is not inhibited thereby. Therefore, it is possible to produce cells efficiently by the continuous culture of Candida utilis under a high cell density. Thus, attention has been paid on Candida utilis as a protein source, and the industrial production of the yeast cell has once been conducted with use of a saccharified liquor of broad-leaved trees or a sulfite waste liquor containing a large amount of pentoses. The yeast, Candida utilis, as well as Saccharomyces cerevisiae and Saccharomyces fragilis has been authorized by U.S. F
REFERENCES:
patent: 5436136 (1995-07-01), Hinnen et al.
Rose et al. Methods in Enzymology, vol. 194, pp.199-206, 1991.
Hsu et al. Journal of Bacteriology, vol. 154(3), pp. 1033-1039, Jun. 1991.
Rothstein, R. Methods in Enzymology, vol. 194, pp.281-301, 1991.
Dehoux et al. European Journal of Biochemistry, vol. 213, pp. 841-848,1993.
Kawai et al. Journal of Bacteriology, vol. 173(1), pp. 254-262, 1992.
Lopes et al. Gene, vol. 79, pp. 199-206, 1989.
Russel et al. Biochemica et Biosphysica Acta, vol. 1008, pp. 243-246, 1989.
Becker et al. Methods in Enzymology, vol. 194, pp. 281-301, 1991.
Kajiwara Susumu
Kondo Keiji
Misawa Norihiko
Ketter James
Kirin Beer Kabushiki Kaisha
Yucel Irem
LandOfFree
Transformation systems for the yeast candida utilis and the expr does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Transformation systems for the yeast candida utilis and the expr, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Transformation systems for the yeast candida utilis and the expr will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1456741