Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1998-03-09
1999-09-14
Degen, Nancy
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 692, 435 693, 435 694, 435 695, 435 696, 435 699, 4353201, 435440, 536 231, 536 241, C12P 2102, C07H 2104, C12N 1563
Patent
active
059521953
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to the field of biotechnology, in particular to an improvement made to the production of a polypeptide of commercial or therapeutic interest in yeast and, in particular, in Saccharomyces cerevisiae. It also relates, firstly, to new nucleic acid fragments isolated from the genomic DNA of Saccharomyces cerevisiae and having a transcriptional promoter activity and, secondly, expression cassettes, expression vectors and host cells containing them as well as their use for the production of polypeptides of interest.
The yeast Saccharomyces cerevisiae is considered as one of the preferred hosts for the production of recombinant proteins for numerous reasons. On the one hand, this organism is nonpathogenic and it is commonly used in the food industry. On the other hand, it can be cultured on a large scale and at high density in a relatively inexpensive medium and can be easily adapted to an industrial environment. Furthermore, it has been particularly studied, such that many data relating to its genetics and its physiology are available. Finally, it is capable of carrying out certain typically eukaryotic modifications (glycosilation, disulfide bridges and the like).
Although numerous transcriptional promoters which are functional in yeasts have been described in the literature, only some of them have proved effective for the production of polypeptides by the recombinant route. There may be mentioned in particular the promoters of the PGK genes (3-phosphoglycerate kinase; Hitzeman et al., 1983, Science, 219, 620-625), TDH genes encoding GAPDH (Glyceraldehyde phosphate dehydrogenase; Holland and Holland, 1979, J. Biol. Chem., 254, 9839-9845), TEF1 genes (Elongation factor 1; Cottrelle et al., 1985, J. Biol. Chem., 260, 3090-3096), MF.alpha.1 (.alpha. sex pheromone precursor; Inokuchi et al., 1987, Mol. Cell. Biol., 7, 3185-3193) which are considered as strong constitutive promoters or alternatively the regulatable promoter CYCl which is repressed in the presence of glucose (Guarente and Ptashne, 1981, Proc. Natl. Acad. Sci. USA, 78, 2199-2203) or PHO5 which can be regulated by thiamine (Meyhack et al., 1982, EMBO J., 1, 675-680). However, for reasons which are often unexplained, they do not always allow the effective expression of the genes which they control and therefore the production of polypeptides at high levels. In this context, it is always advantageous to be able to have new promoters in order to generate new effective host/vector systems for the production of a large quantity of proteins of interest. Furthermore, having a choice of effective promoters in a given cell also makes it possible to envisage the production of multiple proteins in this same cell (for example several enzymes of the same metabolic chain) while avoiding the problems of recombination between homologous sequences.
In general, a promoter region is situated in the 5' region of the genes and comprises all the elements allowing the transcription of a DNA fragment placed under their control, in particular:
(1) a so-called minimal promoter region comprising the TATA box and the site of initiation of transcription, which determines the position of the site of initiation as well as the basal level of transcription. In Saccharomyces cerevisiae, the length of the minimal promoter region is relatively variable. Indeed, the exact location of the TATA box varies from one gene to another and may be situated from -40 to -120 nucleotides upstream of the site of the initiation (Chen and Struhl, 1985, EMBO J., 4, 3273-3280)
(2) sequences situated upstream of the TATA box (immediately upstream up to several hundreds of nucleotides) which make it possible to ensure an effective level of transcription either constitutively (relatively constant level of transcription all along the cell cycle, regardless of the conditions of culture) or in a regulatable manner (activation of transcription in the presence of an activator and/or repression in the presence of a repressor). These sequences, subsequently designated modulatory sequenc
Achstetter Tilman
Nacken Valerie
Degen Nancy
Transgene S.A.
LandOfFree
Promoters for expressing proteins of interest in yeast does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Promoters for expressing proteins of interest in yeast, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Promoters for expressing proteins of interest in yeast will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-1509204