240 results about "Stop codon" patented technology

The present invention provides techniques and reagents for read-through of stop codons in mammalian cells. In certain embodiments, the invention provides methods in which a mammalian cell is contacted with a synthesized tRNA so that the tRNA is taken up into the cell at levels that allow read-through of stop codons. Preferably, the synthesized suppressor tRNA is aminoacylated, optionally with an unnatural amino acid. In certain preferred embodiments, the inventive system is utilized to generate proteins containing two or more unnatural amino acids.

In various aspects, the invention provides Brassica juncea plants, seeds, cells, nucleic acid sequences and oils. Edible oil derived from plants of the invention may have significantly higher oleic acid content than other B. juncea plants. In one embodiment, the B. juncea line MJ02-086-3 contains a mutant allele MJ02-086-3/BjFAD2-a at the BjFAD2-a gene locus, having a premature stop codon, so that there are no functional copies of the FAD2 gene in MJ02-086-3 plants. Seeds from MJ02-086-3 plants may for example yield an oil having oleic acid content of greater than about 70% by weight.

The human soluble neuropilin-1 (sNRP) polyadenylation signal (sNRP-poly(A)), situated downstream of the GT splice donor site of intron 12 of the full-length neuropilin-1 gene, also functions as the termination codon for sNRP. This 17 nucleotide sequence efficiently facilitates addition of poly(A) tails to RNAs expressed in cells. The present invention shows that this optimally succinct sequence has similar activity to the SV40 polyadenylation signal that is currently used in expression vectors. By using this shorter dual termination/polyadenylation signal and avoiding the need for large and cumbersome polyadenylation signals, expression vectors may be engineered to carry considerably larger genes.

The invention relates to a Qbeta-2aa phage virus-like particle protein which is prepared according to a method comprising the following steps of: (1) mutating a Qbeta phage CP (coat protein) gene terminator codon into a strong terminator codon TAA from TGA and then cloning the strong terminator codon into a protokaryon expression vector pET28a(+) to obtain a coding CP plasmid pETQB-CP; (2) inserting the nucleotides AAGCTT of coding lysine and leucine at the coding immunodominance determining region position in a Qbeta phage CP extension protein gene, i.e. between the 72th codon and the 73th codon of the Qbeta phage CP extension protein gene, and mutating the CP gene terminator codon into GGA from TGA and then cloning the GGA into a protokaryon expression vector PGEX4T-2 to obtain a codingA1 protein plasmid pGEXQbeta-A1; (3) jointly converting the mutated coding Qbeta phage CP plasmid and the A1 protein plasmid into escherichia coli, and inducing and then expressing to obtain the virus-like particle protein Qbeta-2aa; and (4) coupling a short peptide antigen to Qbeta-2aa phage virus-like particles by using a iso-bifunctional cross-linking agent.

The present invention provides a cyclic RNA preferable for carrying out rotary protein translation in which translation domains other than that of the target protein are sufficiently short and translation efficiency is high, and a method for producing protein that uses this cyclic RNA as template. More specifically, the present invention provides a cyclic RNA that encodes a protein, has a full-length number of bases that is equal to or greater than 102 and is a multiple of 3, has at least one start codon, does not have a stop codon in the same reading frame as the start codon, and does not contain an internal ribosome entry site (IRES). In addition, the present invention provides a method for producing protein in a eukaryotic cell expression system that consists of using the aforementioned cyclic RNA as template and expressing a protein encoded by that cyclic RNA.

The invention belongs to the field of animal gene engineering, and in particular relates to a synthetic fusion gene for expressing mycoplasma hyopneumoniae and application. The fusion gene is characterized in that a mycoplasma hyopneumoniae P97R1 gene is connected in series with a P36 gene through a Linker, the termination codon of the P36 gene is deleted, the P46 gene with signal peptide removed is fused at the C end of the P36 gene through Linker, and then the fusion gene P97R1-P36-P46 is obtained, wherein the nucleotide sequence of the fusion gene is shown as SEQ ID NO: 1. The fusion gene is included in a prokaryotic expression plasmid, and escherichia coli BL21/pET30a-P97R1-Linker-P36-Linker-P46 containing the plasmid is collected in CCTCC with the collection number CCTCC NO: M2014269. The invention further discloses immune efficacy evaluation of the fusion gene and application of the fusion gene in a novel vaccine.

The invention discloses a neutral endoglucanase as well as an encoding gene and an application thereof. An H31 gene sequence is an intact open reading frame (ORF), wherein the open reading frame begins from an initiation codon ATG and is terminated by a termination codon TAG, and comprises 762 nucleotides; the nucleotide sequence is shown in SEQ ID No:1; an amino acid sequence of protein encoded by the neutral endoglucanase H31 gene is shown in SEQ ID No.2. The invention further discloses an application of the neutral endoglucanase H31 in industries such as washing and papermaking.

The invention discloses a manually-combined Newcastle diseases virus F gene and a recombining expression vector and an application thereof. Codons of NDVF gene OFR are all replaced into chicken bias codons, EcoR V restriction sites and Kozak sequences are added upstream, Xbal I restriction sites and TGA stop codons are added downstream, and downstream bases A of the stop codons are changed into G, obtained genes (SEQ, ID NO: 2) can be efficiently expressed in eukaryotic cells, and the expression efficiency of the obtained genes is higher than that of wild type genes. Expressed genes have immunogenicity and biologic activity of natural protein of Newcastle diseases virus, can stimulate chicken to generate body fluid stronger immunologic response and stronger cell immunologic response than the wild F gene DNA vaccine so as to enable immunized chicken to obtain 100 percent of resistance to fatal attack of high-pathogenicity Newcastle strains, and can effectively inhibit the propagation of the virus in the chicken.

The invention discloses an alkaline endoglucanase gene. The gene sequence is a complete open reading frame, which begins with ATG start codon and concludes with TAA stop codon. The whole sequence comprises 2502 nucleic acids. The invention also discloses the application of the recombinant alkaline endoglucanase in detergent industry etc. The recombinant alkaline endoglucanase has good resistance for SDS, which is preserved in the detergent with 0.2 percent final concentration for ten minutes without significant activity loss of the residue enzyme. The recombinant alkaline endoglucanase suits for the detergent and textile industries etc.

The present invention relates to an isolated major gene GS3 which regulates grain weight and grain length in the rice and the cloning of said gene. The DNA sequence of GS3 gene is as shown in SEQ ID NO. 1 and is 7883 bp in length. GS3 gene comprises 5 exons and encodes 232 amino acids. It is predicted based on bioinformatics analysis that said protein contains conserved domains including a PEBP-like domain, a transmembrane domain, a cysteine-rich domain of TNFR/NGFR and a VWFC domain. cDNA sequence of said gene is as shown in SEQ ID NO. 2. By sequence alignment between three large grain species and 3 small grain species of rice, it is revealed there is only one common single nucleotide mutation in a 7.9-kb region between the two different grain-length groups. Said nucleotide mutation is located at the second exon of the GS3 gene, in which a cysteine codon (TGC) in the small-grain group is mutated to a termination codon (TGA) in the large-grain group. This mutation causes a premature termination in the large-grain group, which leads to a 178-amino acids truncation (including part of the PEBP-like domain and all the other three conserved domains). The present invention also provides methods of producing transgenic plants comprising sequences disclosed herein.

The invention provides a method for synthesizing an LfcinB15-Mag12 encoding gene and expressing the same in colon bacillus. Active gene fragments enconding LfcinB 1-15 amino acid and active gene fragments of Magainin from 1 to 12 are designed according to E. coli codon preference, a recombinant expression vector pET-32a-Lfcin B15-Mag12 is constructed by an expression vector pET-32a, codon encoding 6 histidine labels exists before multiple cloning sites of pET-32a, and stop codon TAG and TAA are added. The method successfully expresses the LfcinB15-Mag12 in the colon bacillus by a gene engineering method, establishes basis of theory and practice for scale production of hybrid peptide and other noble peptides by the gene engineering method, and is significant for establishing peptide gene engineering strains and commonness technology of antibacterial peptide research.

The invention is the gene engineering preparation polypeptide remedy technology field of biology technology. The aid is to build a double function fusion albumen gene of rhTPO/SCF, express and prepare with more biology activityú¼less side-effect rhTPO/SCF in insect cell. The gene of rhTPO/SCF includes 1-157 amino acids coding sequence with TPO, 16 amino acids continual peptide coding sequence, 1-145 amino acids coding sequence of SCF and codoní»s DNA fragments. By taking the rhTPO/SCF into TF1 cells experiment, which preparation from the infect recombination virus growing in adherence S19 cell, it mensurates that the specific activity is 2.0 í‡10 to the power 5 units/mg; by Mo7e cells experiment, the specific activity is 8.3í‡10 to the power 5 units/mg. It can be used to cure low blood platelet cause by radiotherapy, chemotherapy, bone marrow transplantationíú

The present invention provides recombinant expression cassettes comprising a fungal 3′ termination sequence which is functional in a plant. The recombinant expression cassettes comprise a plant promoter operably linked to a coding sequence having a stop codon, and the fungal 3′ termination sequence. The fungal 3′ termination sequence is heterologous to the coding sequence. The fungal 3′ termination sequence comprises structural features including a cleavage site, a positioning element, and an upstream element. The present invention also comprises methods for construction of the plant expression cassettes and introducing the cassettes into plant cells.

The invention discloses fusion protein capable of improving dammarenediol conversion efficiency, a construction method and application. The construction steps comprise: excising first 138 bases at 5' end of arabidopsis thaliana cytochrome-NADPH-reductase 1 gene AtCPR1, so as to obtain a sequence shown as SEQ ID NO. 2; (2) removing a termination codon TAA at 3' end of ginseng protopanaxadiol synthase gene PPDS with a sequence shown as SEQ ID NO. 3, and connecting with the base sequence which at 5' end of the sequence shown as SEQ ID NO. 2 and used for coding polypeptide GSTSSGSG, so as to construct a gene member of fusion protein; and (3) connecting the gene member of the fusion protein with saccharomyces cerevisiae cell endogenous promoter and terminator, so as to construct a fusion protein gene expression cassette, and transforming into saccharomyces cerevisiae cell for expression. The fusion protein constructed by the method is capable of improving the conversion rate of dammarenediol to protopanaxadiol.

The invention discloses alkaline xylanase as well as a coding gene and an application thereof and belongs to the technical field of biology. The gene sequence of WMN-3 is a complete ORF (open readingframe), wherein the ORF starts with start codon ATG, stops with stop codon TAA and totally contains 1476 nucleotides, and the nucleotide sequence is shown as SEQ ID NO:1. An amino acid sequence of a protein coded by a gene of alkaline xylanase WMN-3 is shown as SEQ ID NO:2. The invention further discloses the application of the alkaline xylanase WMN-3 to industries such as papermaking and the like.

The invention discloses a method for cloning the complete sequence of goat MC4R gene coding region, comprising the following steps: A1, extracting total RNA in goat eye muscle tissue, and then reverse-transcribing the total RNA into cDNA; A2, designing primers and PCR Reaction: The upstream primer is designed in the upstream region of the start codon, and the downstream primer is designed in the downstream region of the stop codon; the designed primers are: upstream primer P1: 5′-CTCTCCGCCGCCTAACTTTCGTTT-3′ downstream primer P2: 5′-GTGCCTATAACCTGTGATGTGGAT- 3'. A3, recovery and purification of PCR products; A4, cloning. The invention provides a simple and fast method for obtaining the complete coding region sequence of goat MC4R gene, fills in the blank of the gene in goat, and lays the foundation for further research on the gene of goat.

The invention discloses an alpha-L-arabinofuranosidase capable of improving the filtration performance of wort, and belongs to the technical field of bioengineering. According to the present invention, the gene of alpha-L-arabinofuranosidase is modified, wherein the intron, the signal peptide and the stop codon sequence are removed, the sequence of the modified alpha-L-arabinofuranosidase gene isrepresented by SEQ ID NO.4, and the corresponding amino acid sequence is represented by SEQ ID NO.2; the sequence of the modified alpha-L-arabinofuranosidase gene is linked to an expression vector pPIZ[alpha]A to construct a recombinant expression vector pPIZ[alpha]A-QAnabfA, and expression is performed in Pichia pastoris X33 to obtain recombinant alpha-L-arabinofuranosidase; and with the application of the recombinant alpha-L-arabinofuranosidase in the production of wort, the filtration speed of the wort can be improved.