Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1996-12-16
1999-12-07
Brusca, John S.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 914, 4352523, 43525411, 4353201, 435325, 435419, 536 231, 536 241, 536 245, C12P 2102, C12N 1511, C12N 1563, C12N 504
Patent
active
059981679
DESCRIPTION:
BRIEF SUMMARY
This invention relates to expression control polynucleotides derived from spliceosomal protein gene promoters.
It is already well known that organisms such as plants or animals with novel characteristics can be produced by introducing genes or DNA sequences from the same or a related organism or by introducing genes or DNA sequences from other organisms. The key to production of novel phenotypes is the active expression of at least a part of the introduced DNA sequence. Generally expression of the introduced DNA sequence will occur only in the presence of an expression control polynucleotide, such as a promoter, which is compatible with the host organism. Promoters are nucleic acid sequences which are currently believed to regulate the expression of a gene by facilitating the binding of proteins required for transcription, such as RNA polymerase, to a portion of the nucleic acid sequence upstream of the gene. Thus for protein coding genes the DNA is transcribed into mRNA (messenger RNA) which is then relocated to the cytoplasm where it is available for translation into polypeptide.
Whereas promoters are generally disposed upstream of the genes they regulate and are thought to act by providing a binding site for an RNA polymerase transcription complex, another form of expression control polynucleotide known as an "enhancer" sequence can control the expression of a gene usually without regard to its relative position or orientation.
Promoters can vary in levels of expression induction and in their expression patterns. Some promoters are active in a tissue-specific or developmental stage-specific manner while other promoters are active constantly to continually drive expression of the gene they control. Such constantly active promoters are called constitutive promoters. The promoters of this latter class most widely used in genetic engineering are the Cauliflower Mosaic Virus (CaMV) 35S RNA promoter, nopaline synthetase (nos) or octopine synthetase (ocs) promoters. Constitutive promoters induce gene expression at a relatively constant rate.
Although the above conventional promoters generally drive expression at high levels, many have the disadvantage that they are derived from plant infectious agents; the CaMV 35S RNA promoter is derived from a plant virus and the other promoters mentioned are derived from strains of Agrobacterlium, soil-borne infectious bacteria. The source of these promoters is a cause of concern in transgenic plant production. In addition, detailed analysis of expression patterns of CaMV 35S promoter, have shown that its levels of expression can vary greatly among different plant tissues to the level where it is inactive in some tissues and is therefore, no longer constitutive for such tissue. For many biotechnological objectives, constitutive expression in all cells and tissues would be of great advantage.
Splicesomal proteins are believed to be present in virtually all eukaryotic cells and are involved in the phenomenon of pre-mRNA splicing which removes introns (non-coding regions) from RNA transcripts before protein production.
The present invention seeks to provide a promoter (or other expression control polynucleotide) which is not derived from an infectious agent and which is suitable for use in the control of expression of recombinant genes in the construction of transgenic organisms such as plants and animals.
The present invention also seeks to provide a promoter (or other expression control polynucleotide) which is likely to be active throughout all or most cells of the organism.
According to the present invention there is provided an expression control polynucleotide at least partially derived from a spliceosomal protein gene promoter.
The expression control polynucleotide of the invention is capable of controlling the expression of a second polynucleotide (preferably comprising a polypeptide-encoding sequence) operably linked thereto.
RNA sequences which do not code for protein can also be expressed eg ribozymes or anti-sense RNA.
The term "expression control polynucleotide" a
REFERENCES:
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G. Biamonti, et al., Two Homologous Genes, Originated by Duplication, Encode the Human HNRNP Proteins A2 and A1, vol. 22, No. 11, Jun. 11, 1994, pp. 1996-2002 Nucl. Acids. Res.
G. Biamonti, et al, Human HNRNP Protein A1 Gene Expression Structural and Functional Characterization of the Promoter, vol. 230, May 3, 1993, pp. 77-89 J. Mol. Biol.
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H. Kulesza, et al., Detection of a Plant Protein Analagous to the Yeast Spliceosomal Protein PRP8. Febs Letters, vol.318, No. 1, Feb. 1993, pp. 4-6.
T. Kiss, et al. Molecular Analysis of a U3 RNA Gene Locus in Tomato; Transcription Signals, The Coding Region, Expression in Transgenic Tobacco Plants and Tandemly Repeated Pseudogenes, Nucleic Acids Research, vol. 18, No. 8, 1990, pp. 1941-1949.
D. Edoh, et al. Activity of U-SNRNA Genes with Modified Placement of Promoter Elements in Transfected Protoplasts and Stably Transformed Tobacco, Nucleic Acids Research, vol. 21, No. 7, 1993, pp. 1533-1540.
Brown John Willaim Slessor
Clark Gillian Patricia
Simpson Gordon Grant
Brusca John S.
Scottish Crop Research Institute
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