101 results about "Plastid" patented technology

Disclosed are the uses of specific genes of the mevalonate and isoprenoid biosynthetic pathways, and of inactive gene sites (the pseudogene) to (1) enhance biosynthesis of isopentenyl diphosphate, dimethylallyl diphosphate and isoprenoid pathway derived products in the plastids of transgenic plants and microalgae, (2) create novel antibiotic resistant transgenic plants and microalgae, and (3) create a novel selection system and / or targeting sites for mediating the insertion of genetic material into plant and microalgae plastids. The specific polynucleotides to be used, solely or in any combination thereof, are publicly available from GeneBank and contain open reading frames having sequences that upon expression will produce active proteins with the following enzyme activities: (a) acetoacetyl CoA thiolase (EC 2.3.1.9), (b) 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) synthase (EC 4.1.3.5), (c) HMG-CoA reductase (EC 1.1.1.34), (d) mevalonate kinase (EC 2.7.1.36), (e) phosphomevalonate kinase (EC 2.7.4.2), (f) mevalonate diphosphate decarboxylase (EC 4.1.1.33), (g) isopentenyl diphosphate (IPP) isomerase (EC 5.3.3.2), and (h) phytoene synthase (EC 2.5.1.32).

Provided are methods for increasing the production of protein in a plant cell by transforming plastids of plant cells with a construct comprising a promoter functional in a plant plastid, a ribosome binding site, DNA sequence of interest and a transcription termination region, and growing plant cells comprising the transformed plastids under conditions wherein the DNA encoding sequence is transcribed in the plastid. Also provided are methods for increasing protein production by fusing a coding sequence to a gene of interest to a secondary protein for cleavage or targetting of the protein of interest within the plastid, whereby high levels of expression of protein is achieved.

Transgenic chloroplast technology could provide a viable solution to the production of Insulin-like Growth Factor I (IGF-I), Human Serum Albumin (HSA), or interferons (IFN) because of hyper-expression capabilities, ability to fold and process eukaryotic proteins with disulfide bridges (thereby eliminating the need for expensive post-purification processing). Tobacco is an ideal choice because of its large biomass, ease of scale-up (million seeds per plant), genetic manipulation and impending need to explore alternate uses for this hazardous crop. Therefore, all three human proteins will be expressed as follows: a) Develop recombinant DNA vectors for enhanced expression via tobacco chloroplast genomes b) generate transgenic plants c) characterize transgenic expression of proteins or fusion proteins using molecular and biochemical methods d) large scale purification of therapeutic proteins from transgenic tobacco and comparison of current purification / processing methods in E. coli or yeast e) Characterization and comparison of therapeutic proteins (yield, purity, functionality) produced in yeast or E. coli with transgenic tobacco f) animal testing and pre-clinical trials for effectiveness of the therapeutic proteins. Mass production of affordable vaccines can be achieved by genetically engineering plants to produce recombinant proteins that are candidate vaccine antigens. The B subunits of Enteroxigenic E. coli (LTB) and cholera toxin of Vibrio cholerae (CTB) are examples of such antigens. When the native LTB gene was expressed via the tobacco nuclear genome, LTB accumulated at levels less than 0.01% of the total soluble leaf protein. Production of effective levels of LTB in plants, required extensive codon modification. Amplification of an unmodified CTB coding sequence in chloroplasts, up to 10,000 copies per cell, resulted in the accumulation of up to 4.1% of total soluble tobacco leaf protein as oligomers (about 410 fold higher expression levels than that of the unmodified LTB gene). PCR and Southern blot analyses confirmed stable integration of the CTB gene into the chloroplast genome. Western blot analysis showed that chloroplast synthesized CTB assembled into oligomers and was antigenically identical to purified native CTB. Also, GM1-ganglioside binding assays confirmed that chloroplast synthesized CTB binds to the intestinal membrane receptor of cholera toxin, indicating correct folding and disulfide bond formation within the chloroplast. In contrast to stunted nuclear transgenic plants, chloroplast transgenic plants were morphologically indistinguishable from untransformed plants, when CTB was constitutively expressed. The introduced gene was stably inherited in the subsequent generation as confirmed by PCR and Southern blot analyses. Incrased production of an efficient transmucosal carrier molecule and delivery system, like CTB, in transgenic chloroplasts makes plant based oral vaccines and fusion proteins with CTB needing oral administration a much more practical approach.

The invention discloses a method for increasing the crystallization ratio of crystallized malt, and belongs to the technical field of beer brewing. According to the method, barley serving as a raw material is malted, saccharified, baked and charred to obtain the full crystallized malt with glass plastids inside. In the malting process, steeping temperature (14-15 DEG C) is slightly increased, water is repeatedly supplemented in the germination process, and the water content of the green malt is increased and about 42%-44%. In the proteolysis and saccharification process, in order to ensure uniformity of the water content and the quality of the malt, about 200g of green malt is directly soaked in 1L of water and saccharified. At present, the crystallization ratio of the crystallized malt can be only about 20%, but the crystallization ratio of the crystallized malt prepared by the method can reach 80% or more.

The invention discloses application of a diatom UPA (universal plastid amplicon) gene in judging whether the drowned dead really die of drowning. The invention provides a pair of primer pairs for detecting the diatom UPA gene and further provides a diatom UPA gene analyzing method. The analyzing method comprises the steps of extracting diatom nucleic acid, utilizing the primer pair shown in the right of claim 2 to amplify the UPA gene in the nucleic acid, utilizing an amplified product to prepare a sample to be tested and performing nucleic acid analysis on the sample to be tested. Furthermore, a used probe is 5''-CGGAGGCGTACAAAG-3'', the 5'' end of the probe is marked by an FAM fluorescent reporter group, and the 3'' end of the probe is marked by an NFQ-MGC MGB fluorescent quencher group. The invention further provides application of the diatom UPA gene analyzing method in legal medical expert detection. The application of the diatom UPA gene analyzing method in legal medical expert detection comprises the steps of extracting the diatom nucleic acid in organs of the drowned dead, amplifying the diatom nucleic acid through a PCR technology, then detecting and comparing the diatom nucleic acid with diatom nucleic acid in a water sample. Through comparison of relevance ratio and a sequencing result of the diatom UPA gene, whether the drowned dead really die of the drowning can be judged. Meanwhile, the application of the diatom UPA gene analyzing method in legal medical expert detection is beneficial to finding out a real drowned area.

This disclosure concerns the systems, methods, and kits for the in vitro synthesis of biological macromolecules in a reaction utilizing cell lysates containing plastids, mitochondria and / or chloroplasts, wherein creatine phosphate and creatine kinase are not added to the reaction to provide artificial energy regeneration.

A process for producing multicellular plants, plant organs or plant tissues transformed on their plastome by the following steps is provided: (a) altering or disrupting the function of a gene in a plastid genome for producing a selectable or recognizable phenotype; (b) separating or selecting plants or cells having plastids expressing said phenotype; (c) transforming said plastid genome of said separated or selected plant, plant organ or plant tissue with at least one transformation vector having a restoring sequence capable of restoring said function; and (d) separating or selecting said transformed plant, plant organ or plant tissue having plastids expressing said restored function.

This invention provides cyanobacteria as an alternative source of ahas and pds genes for plant transformations and for selectable markers. In particular, it provides for cyanobacteria, for example, Synechocystis, as a source of genes encoding herbicide insensitive proteins, and elements of genes for control of expression in plastids. Nucleic acid fragments, both the acetolactate synthase (ahas) large subunit and the ahas small subunit, were found to provide herbicide resistance. Also, the present invention provides novel Synechocystis mutant phytoene desaturase (PDS) gene conferring resistance to 4′-fluoro-6[(alpha,alpha,alpha,-trifluoro-m-tolyl)oxy]-picolinamide, a bleaching herbicide. The present invention provides improvements to method involving cyanobacteria for the screening of compounds, including a new high-through-put protocol that is a rapid and cost effective way to identify target site genes.

This invention provides a novel method to confer disease resistance to plants. Plant plastids are transformed using a plastid vector which contains heterologous DNA sequences coding for a cytotoxic antimicrobial peptide. Transgenic plants are capable of fighting off phytopathogenic bacterial infection.

Production of human serum albumin (HSA) in prokaryotic systems has not been successful to date because HSA is highly susceptible to proteolytic degradation. Production in plants has not yielded enough protein to be cost-effective. The instant invention overcomes this by producing HSA in plant plastids at high levels.