281 results about "Synthetic gene" patented technology

The present invention is drawn to methods for designing synthetic nucleotide sequences encoding polypeptides of interest. The methods involve organizing a database of sequences as a set of N-length oligomer sequences and compiling a list of probability scores for each N-length sequence. The probability scores are used to substitute one or more higher-scoring sequences into the parent nucleotide sequence to generate an optimized sequence. The nucleotide sequence of interest may be further optimized by removing either or both of unintended open reading frames or undesired short DNA elements, and/or substituting oligomer sequences to achieve a specific G:C content. These methods may be used for optimizing expression of heterologous genes in any organism, particularly in plants. The method generates synthetic sequences with a composition similar to that of a target database. These synthetic sequences may be used, for example, for regulating pesticidal activity or herbicide resistance in organisms, particularly plants or plant cells.

This invention is directed to preparation and expression of synthetic genes encoding polypeptides containing protective epitopes of botulinum neurotoxin (BoNT). The invention is also directed to production of immunogenic peptides encoded by the synthetic genes, as weel as recovery and purification of the immunogenic peptides from recombinant organisms. The invention is also directed to methods of vaccination against botulism using the expressed peptides.

This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an α-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the α-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.

Methods for making synthetic gene clusters are described.

Disclosed herein is a method for synthesizing a desired nucleic acid sequence. The method comprises dividing the desired sequence into a plurality of partially overlapping segments; optimizing the melting temperatures of the overlapping regions of each segment to disfavor hybridization to the overlapping segments which are non-adjacent in the desired sequence; allowing the overlapping regions of single stranded segments which are adjacent to one another in the desired sequence to hybridize to one another under conditions which disfavor hybridization of non-adjacent segments; and filling in, ligating, or repairing the gaps between the overlapping regions, thereby forming a double-stranded DNA with the desired sequence. Also disclosed is a method for preventing errors in the synthesis of the nucleic acid sequence.

The invention discloses a bacterial strain for producing farnesene and an application of the bacterial strain, belonging to the field of synthetic biology. The bacterial strain for producing the farnesene contains related genes for synthesizing the farnesene through a mevalonic acid pathway and codon optimization; and the sequence of a farnesene synthetic gene afs optimized by codons is as shown by SEQ ID NO.1. The bacterial strain can be used for producing the farnesene, and a seed solution of the bacterial strain is inoculated into a culture medium containing a carbon source to carry out prokaryotic expression to obtain the farnesene. The gene for synthesizing the farnesene is subjected to codon optimization or the farnesene is synthesized by using the mevalonic acid pathway to ensure that each protein is closer to a ratio of AtoB:ERG13:tHMG1:ERG12:ERG8:MVD1:Idi:IspA:AFS=1:10:2:5:5:2:5:2:2, so that the production of the farnesene can be further promoted. By adopting the bacterial strain, the output of the farnesene is greatly improved to be more than 1g/L.

The present application relates to somatic hypermutation (SHM) systems and synthetic genes. Synthetic genes can be designed using computer-based approaches to increase or decrease susceptibility of a polynucleotide to somatic hypermutation. Genes of interest are inserted into the vectors and subjected to activation-induced cytidine deaminase to induce somatic hypermutation. Proteins or portions thereof encoded by the modified genes can be introduced into a SHM system for somatic hypermutation and proteins or portions thereof exhibiting a desired phenotype or function can be isolated for in vitro or in vivo diagnostic or therapeutic uses.

A new approach in the field of plant gums is described which presents a new solution to the production of hydroxyproline(Hyp)-rich glycoproteins (HRGPs), repetitive proline-rich proteins (RPRPs) and arabino-galactan proteins (AGPs). The expression of synthetic genes designed from repetitive peptide sequences of such glycoproteins, including the peptide sequences of gum arabic glycoprotein (GAGP), is taught in host cells, including plant host cells.

A new approach in the field of plant gums is described which presents a new solution to the production of hydroxyproline(Hyp)-rich glycoproteins (HRGPs), repetitive proline-rich proteins (RPRPs) and arabinogalactan-proteins (AGPs). The expression of synthetic genes designed from repetitive peptide sequences of such glycoproteins, including the peptide sequences of gum arabic glycoprotein (GAGP), is taught in host cells, including plant host cells.

The invention discloses an engineering bacterium through anthracene nucleus antibiotic and appliance, which is characterized by the following: inserting or replacing restriction enzymes site of dnmV gene nucleic acid order of sky blue rufus streptomycete with antibiotic against property gene and surface cerubidin synthetic gene; blocking-up engineering bacterium of dnmV gene; or transforming expressing carrier of anthracene nucleus antibiotic synthetic gene to lead streptomycin (S.lividans)TK24; extracting expressing plasmid from S.lividans TK24; leading-in sky blue rufus streptomycete gene only with antibiotic against property gene to block dnmV gene; blocking-up abrupt change bacterium. This invention can produce cerubidin and/or analogue, such as anthracene nucleus antibiotic.

Synthetic polynucleotides encoding human carcinoembryonic antigen (CEA) are provided, the synthetic polynucleotides being codon-optimized for expression in a human cellular environment. The gene encoding CEA is commonly associated with the development of human carcinomas. The present invention provides compositions and methods to elicit or enhance immunity to the protein product expressed by the CEA tumor-associated antigen, wherein aberrant CEA expression is associated with a carcinoma or its development. This invention specifically provides adenoviral vector and plasmid constructs carrying codon-optimed human CEA and discloses their use in vaccines and pharmaceutical compositions for preventing and treating cancer.

The invention is a method for producing lactic acid for a substrate by glycerin, belonging to the field of biochemical technology. The method comprises the steps that: strains containing glycerin use genes and lactic acid synthetic genes are inoculated to a glycerin-containing seed culture medium; seed liquid is added to an initial fermentation medium containing 30-80g/L glycerin for anaerobic fermentation or micro-aerobic fermentation under 37 DEG C to 55 DEG C; the pH value is adjusted by fed batch 3-4M aqueous slkali or portion-wise addition of solid CaCO<3>, when the production strength of the lactic acid is less than 0.3-0.5g/(1h); the glycerin is stopped fed batching, the fermentation is continued until the residual glycerin concentration decreased to 5g/L. The method has the advantages that: the by product glycerin in the biodiesel production process is reasonably used, and transferred to high-added-value lactic acid; the method can also be used for coupling production of biodiesel and lactic acid, by which high-added-value biodiesel and lactic acid are produced with low-cost raw material, the utilization rate of raw material is improved, and the production cost is lowered.

The invention provides a transcription factor Csa5G157230 participating in regulation of synthesis of cucumber cucurbitacine C and application thereof. The transcription factor is one of two bHLH transcription factors (Csa5G157230 and Csa5G156220) which are found in cucumber genome firstly and used for controlling synthesis of bitter taste; the two bHLH transcription factors are used for controlling the forming of the bitter taste in fruits and leaves respectively; simple crossing of yeast, a gel tissue system and instantaneous expression of tobacco prove that the two transcription factors can be combined to a bitter taste synthesis promoter region and used for activating transcription of a synthesis gene; meanwhile, the bitter taste characteristics of cucumber plants can be restored by expressing the corresponding transcription factors in the leaves or fruits of the cucumber free of bitter taste abundantly. The invention further discloses a molecular mechanism for forming bitter taste of cucumber and provides theoretical basis for breeding cucumber free of bitter taste and molecular assisted breeding goals.

The invention discloses a biosynthetic gene cluster of paquete amide and an application thereof. The biosynthetic gene cluster of paquete amide comes from (Streptomyces pactum)SCSIO 02999, and the cluster comprises five genes including heterozygous polyketone/ nonribosomal peptide synthetases genes ptmA, FAD dependent redox enzyme genes ptmB1, phytoene dehydrogenase genes ptmB2, cyclase genes ptmC and hydroxylase genes ptmD. According to the biosynthetic gene cluster of paquete amide and the application thereof, information in genes and proteins related to biosynthesis of the paquete amide provides theoretical foundations and materials for conducting genetic modification on the biosynthesis of multi-ring tetramate macrocylic lactam family. By conducting genetic modifications on the biological synthetic genes, 6 new structures antineoplastic paquete amide compounds pactamide A-F are obtained, and thus effective compound entities are provided for research and development of antineoplastic drugs.

The present invention belongs to the field of antibiotic gene engineering, and is especially the process of cloning nicomycin biological synthetic gene sanU from one strain of Streptomyces ansochromogenes 7100 CGMCC 4.321; connecting the synthetic gene sanU with promoter of melanin biosynthetic structure gene, cloning onto the polycloning site of integrative single-copy plamid, transforming wild Streptomyces ansochromogenes 7100 CGMCC 4.321 protoplast to obtain nicomycin X component of the recombinant engineering bacterium in the yield 2 times higher than wild strain. The high-yield engineering bacterium may be used in preparing antiseptic medicines.

The invention discloses an astaxanthin synthetic gene recombinant expression plasmid pET-Ast with a base sequence as shown in SEQ ID NO: 1. The invention also discloses a preparation method of the recombinant expression plasmid pET-Ast, an application of the recombinant plasmid pET-Ast to the detection of astaxanthin synthetase activity of a target receptor, and an application method. After the astaxanthin synthetic gene recombinant expression plasmid pET-Ast provided by the invention is introduced to an Escherichia coli BL21 strain, the accumulation of astaxanthin in a cell body of Escherichia coli is successfully realized, so that an astaxanthin production strain can be accurately and rapidly established. The preparation method of the astaxanthin synthetic gene recombinant expression plasmid pET-Ast is simple and convenient, easy to operate and low in cost. By using the method disclosed by the invention, a gene sequence to be detected in the target receptor can be replaced with a related gene sequence in the recombinant expression plasmid pET-Ast through digestion and connection, and then, the function of a gene to be detected in the target receptor can be simply and effectively detected and verified through detecting whether the astaxanthin is accumulated in a host cell body.

The invention provides a process for synthesizing genes and other long double stranded polynucleotides by assembling very short oligonucleotides into partly double stranded polynucleotides, and then connecting these partly double stranded polynucleotide subassemblies with linkers comprised of very short oligonucleotides. In one embodiment, the correct order of the polynucleotide subassemblies is coded in overhangs present at each end of the partly double stranded polynucleotide subassemblies. Linkers having a sequence complimentary to the combined overhangs connect adjacent subassemblies, which are then ligated together. In one preferred embodiment the oligos are six bases long, for which there are only 4096 different possible sequence permutations. A complete library of oligos of this size and scale can be cost-effectively synthesized and quality controlled, avoiding the typical errors and yield issues associated with phosphoramidite synthesis of longer oligos. Furthermore, the limited oligo library size supports development of a laboratory-scale gene synthesis machine.