43 results about "Inverted Repeat Sequences" patented technology

The invention provides an insect-resistant gene Cry1Ab-t and an encoding protein thereof. In the gene, by using a plant bias codon and reducing a sequence rich in AT, an inverted repeat sequence and a subsequence-containing undefined eucaryotic deoxyribose nucleic acid (DNA) sequence in the original DNA sequence on the premise of keeping part of amino acid sequences of a Bacillus thuringHansis (Bt) protein Cry1Ab unchanged, a new Cry1Ab-t gene sequence is synthesized by artificial modification, then a prokaryotic expression carrier and a plant expression carrier are constructed for the gene, and a corresponding host cell is converted. In vitro experiments prove that a synthesized Bt toxoprotein has an obvious insecticidal effect on corn borers. The insect-resistant gene Cry1Ab-t and a target protein expressed by the gene can be steadily and efficiently expressed in corns, so that the gene can be used for culturing Cry1Ab-t transgene insect-resistant corns. The artificially synthesized Bt insect-resistant gene Cry1Ab-t can be also used for improving the insect resistance of other crops, fruit trees, vegetables and the like.

The invention belongs to the field of biotechnology, and provides an RNAi vector pGEM-PCTRi capable of mediating the resistance to tobacco viral diseases in China. Analyzing the genome characteristics of main virus groups in China, which comprise Potato Virus Y (PVY), Cucumber Mosaic Virus (CMV) and Tobacco Mosaic Virus (TMV) and then selecting three segments of conserved effective sequences capable of inducing RNAi in PVY CP, CMV RNA1 and TMV CP; and integrating the three segments in a tabling manner and constructing an inverted repeat sequence so as to obtain an RNAi vector pGEM-PCTRi. Massproduction of dsRNA can be realized in escherichia coli by utilizing the vector, and the resistance of tobacco to PVY, CMV and TMV can be induced to generate after the tobacco treatment. The invention has the potential of being applicable to the prevention of viral diseases for tobacco farmland production, and endowed with the advantages of broad spectrum, high efficiency, no reference to transgenosis, and the like.

The invention belongs to plant breeding field, especially the biology engineering breeding to improve the quality of soybean and earthnut. It uses PCR method to clone the specific gene section from the gene of soybean and earthnut. According to gene silence principle, the gene section would be rebuilt into reverse repeating sequence structure, which would be built into the carrier that of promoter and terminator of the specific expression. The gene would be transferred, cultivated, and filtered by farm bacilli induce transformation. The invention could improve the quality of the soybean and the earthnut.

The invention discloses a method for efficiently modifying filamentous fungi and a strain of the modified filamentous fungi. The method is characterized by introducing a multi-gene interference constructing body and a target protein expression constructing body into filamentous fungus cells simultaneously, and then screening to obtain efficiently modified filamentous fungi, wherein the multi-gene interference constructing body comprises promoters, inverted repeat sequences and a terminator, wherein the promoters can initiate transcription in the filamentous fungus cells, the inverted repeat sequences are separated by introns and are connected together by coding sequences of 2-30 target genes in a mosaic manner, and the terminator can terminate transcription in the filamentous fungus cells; the target protein expression constructing body comprises promoters, coding sequences of target proteins and a terminator, wherein the promoters can initiate transcription in the filamentous fungus cells, and the terminator can terminate transcription in the filamentous fungus cells. The method and the strain have the advantages that the modification efficiency of the filamentous fungi is improved; the downstream separation and purification costs of the target proteins are reduced; the filamentous fungus cells modified through one-time transformation can be used for producing food safety level enzyme preparations.

Conventional techniques for preparing a chimera gene having an inverted repeat sequence of a target sequence suffer from complications since a target sequence is inserted into 2 sites on a vector in sense and antisense orientations, and restriction enzyme recognition sequences must be independently provided at an insertion site of a vector and both ends of the target sequence.In this invention, a cassette construct comprising an arbitrary adaptor sequence and an inverted adaptor sequence separated by an arbitrary spacer sequence or a plasmid vector comprising such cassette construct incorporated therein is prepared, a target sequence is ligated to either or both ends of the cassette construct, or a target sequence is inserted into one end of the cassette construct on the plasmid vector, followed by PCR. Thus, a chimera gene having an inverted repeat sequence is prepared.

The invention discloses a DNA fragment for inhibiting the expression of an omega secaline gene in a wheat 1B / 1R translocation line and application thereof. The DNA fragment consists of an omega secaline gene coding region partial sequence, a plant intron sequence and the inverted repeat sequence of the omega secaline gene coding region partial sequence which are connected in turn. An RNAi genetic expression vector containing the DNA fragment is transferred to the wheat 1B / 1R translocation line in a transgenic way, so that the expression of the endogenous omega secaline gene can be inhibited, the adverse effects of omega secaline on the food processing quality of the wheat 1B / 1R translocation line are relieved or eliminated and the food processing quality of the wheat 1B / 1R translocation line is improved.

The invention relates to a method for positioning and integrating transgenosis and an application of the method, and particularly provides a variant loxP element. The method is characterized in that a sequence 'ATAAC' of an inverted repeat sequence in a wild type loxP locus mutates into 'CACCT', and thus the variant loxP element can be obtained. The variant loxP element can greatly improve the integration specificity and integration efficiency of a cre-loxP system, and therefore, efficient local integration of genes can be realized.

The invention relates to construction and application of an E3 region-deleted complete-copy-type recombinant adenovirus 4 carrier system. The system comprises a framework plasmid, a shuttle plasmid and a packaging cell line; the framework plasmid contains 1-26569bp segments on the left side of an adenovirus (AdV) 4 genome, and the adenovirus (AdV) 4 genome has E3 region genes serving as deletion genes; the shuttle plasmid contains cloning sites used for being inserted into a target gene and an inverted repeat sequence on the right side of the adenovirus (AdV) 4 genome; an HEK-293 cell line is adopted as the packaging cell line. Due to the fact that the anthropogenic adenovirus (AdV) 4 genome is safe and reliable, the recombinant adenovirus (AdV) 4 is expected to be applied to gene therapy and a recombinant vaccine in a host which can be infected by the recombinant adenovirus (AdV) 4, that is, the produced recombinant adenovirus (AdV) 4 has safe and wide application.

The invention relates to a method for cultivating cotton resisting verticillium dahliae. The method comprises the step that genes of a sequence shown in the SEQ ID NO.1 and genes of the inverted repeat sequence are guided into a cotton genome. According to the method, verticillium dahliae virulence genes are guided into a cotton plant, so that the genes are transcribed in the cotton plant to generate a double-stranded RNA structure corresponding to the verticillium dahliae virulence genes, Dicer-like protein in the cotton plant cuts the double-stranded RNA structure into siRNA with the size being 20-25nt, and the siRNA can recognize the virulence genes mRNA complementary with the sequence in verticillium dahlia infecting the cotton, degrades the virulence genes and lowers the pathogenicity of verticillium dahlia.

The invention discloses an application of a bombyx mori nuclear polyhedrosis virus (BmNPV) polygene inverted repeat sequence and a bombyx mori lipase-1 gene and specifically an application of the BmNPV polygene inverted repeat sequence and the bombyx mori lipase-1 gene to the construction of a recombinant vector; the vector can express a hairpin RNA (Ribonucleic Acid) for interfering with BmNPV multiple genes and bombyx mori lipase-1; BmNPV is inactivated by the incremental expression of bmlipase-1 (bombyx mori lipase-1) in the intestinal juice of a bombyx mori, and the number of BmNPV which invades intestinal cells of the bombyx mori is reduced; the expressed hairpin RNA can degrade the mRNA (Messenger Ribonucleic Acid) of a BmNPV gene and inhibit the replication and the proliferation of the virus in the Bombyx mori; and after the vector is used for producing a transgenic bombyx mori, the proliferation of BmNPV in the transgenicbombyx mor can be effectively inhibited, and the virus content is reduced, so that the survival rate of the bombyx mori infected with BmNPV is greatly increased.

This invention provides a genetic construct comprising: (a) a DNA cut-and-paste transposon genetic unit which comprises a transposase gene flanked on either side by inverted repeat sequences and operably under the control of a first promoter; and (b) a transgene unit which comprises an expressable transgene, placed operably under the control of a second promoter; wherein the transposon genetic unit and the transgene unit are linked with one of the inverted repeat sequence located in an intron of the transgene.This invention also provides methods of transiently expressing a transgene in a stably transformed eukaryote.This invention further provides methods for obtaining marker-free transgenic plants.

The invention discloses shRNA for inhibiting replication of an SARS-COV-2 virus and an application of the shRNA. The shRNA targets one of E, M and N genes of the SARS-COV-2 virus and comprises any onepair of sequences selected from the following combinations: (a) a reverse repetitive sequence pair consisting of complementary SEQ ID NO: 1 and SEQ ID NO: 2; (b) a reverse repetitive sequence pair consisting of complementary SEQ ID NO: 3 and SEQ ID NO: 4; (c) a reverse repetitive sequence pair consisting of complementary SEQ ID NO: 5 and SEQ ID NO: 6; (d) a reverse repetitive sequence pair consisting of complementary SEQ ID NO: 7 and SEQ ID NO: 8; and (e) a reverse repetitive sequence pair consisting of complementary SEQ ID NO: 9 and SEQ ID NO: 10. The invention also discloses a drug for inhibiting the replication of the SARS-CoV-2 virus in a subject. The drug comprises a vector and a nucleic acid sequence encoding one or more of the shRNAs.

The invention provides a transgenic vector, a method for rapidly constructing an ACE2 humanized animal model by using the transgenic vector, and application of the ACE2 humanized animal model by aiming at research and development of SARS-CoV-2 drugs. The transgenic vector comprises a PiggyBac transposon 5'end inverted repeat sequence (ITR), a CAG promoter, a human ACE2 coding region, a ribosome access site (IRES), firefly luciferase, a dial rat hepatitis post-transcriptional regulatory element (WPR), a polyA site and a 3 'end inverted repeat sequence (ITR), the transgenic vector can efficiently insert human ACE2 and luciferase gene expression cassettes into a mouse genome, and the luciferase and human ACE2 expressed transgenic mice can be rapidly screened through a luciferase living body imaging system.

This invention relates to a method for constructing fusion gene for improving both PVX resistance and PVY resistance of potato. The method comprises: (1) extracting total RNA from PVX-infected and PVY-infected plants, and performing RT-PCR to obtain coat protein (cp) gene of PVX, and replicase gene Nib of PVY; (2) performing PCR to amplify conserved sequences of cp gene and Nib gene; (3) digesting the conserved sequences, and ligating to obtain fusion gene cn; (4) constructing inverted repeat sequence with an intron, a forward cn gene sequence and a backward cn gene sequence to obtain double-resistance gene RNAiCN; (5) connecting the flanking sequences of RNAiCN with promoter and terminator. When the fusion gene is transferred into potato, both PVX resistance and PVY resistance of the potato are improved.

A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid having a Pol I-dependent origin of replication, and an insert including a structured DNA sequence selected from the group consisting of inverted repeat sequence, direct repeat sequence, homopolymeric repeat sequence, eukaryotic origin of replication or eukaryotic promoter enhancer sequence, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction ofreplication. The method also includes modifying the covalently closed circular recombinant molecule such that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, whereby the resultant Pol III-dependent origin of replication covalently closed circular plasmid has improved replication. An antibiotic marker free covalently closed circular recombinantDNA molecule is also provided.

The present invention relates to enhanced Sleeping Beauty-type transposons and methods of transposition. In particular, the invention relates to a polynucleotide comprising a cargo nucleic acid flanked by a left and a right inverted repeat / direct repeat (IR / DR), wherein IR / DRs, having specific sequences, are recognized by a Sleeping Beauty transposase protein and the polynucleotide is capable of integrating into the DNA of a cell. The invention also relates to a kit for transposing a nucleic acid comprising said polynucleotide as well as to further components such as co-factors of transposition capable of depleting a component of the FACT (facilitates chromatin transcription) complex, namely, SSRP1 and / or SUPT16H / SPT16,or an inhibitor of cathepsin selected from the group comprising H,S,V,and L; or a cofactor capable of depleting or inhibiting HSP90; or a factor temporally arresting cells cell cycle in cell cycle phase G0 / G1, G1 / S, or G2 / M; or a factor inhibiting the ubiquitination ofPCNA,or cells wherein these components have been knocked down or inhibited,or the cell cyle arrested in any of said stages. Alternatively or additionally, the kit may comprise as a co-factor of transposition an agent capable of increasing concentration and / or signaling of ATR or a cell wherein concentrationand / or signaling of ATR are increased. The invention further provides methods using said transposon polynucleotide as well as host cells and pharmaceutical compositions. It also relates to use of said co-factors of transposition or specific cells for enhancing transposition efficiencies, e.g., for preparing genetically modified nucleic acids or cells.

The invention discloses a plasmid for constructing an interference vector, a construction method thereof and application thereof. The invention provides a plasmid pFGC-NAP obtained by inserting a DNA presented by sequence 1 in a sequence list between an Stu I enzyme cutting site and an Asc I enzyme cutting site of a plasmid pFGC1008. The interference vector can be obtained by respectively inserting a plus-sense DNA fragment and an anti-sense DNA fragment into a multiple cloning site of the plasmid pFGC-NAP, wherein the plus-sense DNA fragment and the anti-sense DNA fragment are inverted repeat sequences designed for a target gene to be interfered and expressed. The plasmid constructs the interference vector capable of specially expressing a promoter during a seed development period, has high interference efficiency, can control a spatial temporal specificity expression, can be used for a specific gene expressed in silent plant seeds, and improves quality of oil crop seeds or other plant seeds.

The invention provides shRNA of a CXCR4 gene and an application of the shRNA. The shRNA comprises siRNA of the CXCR4 gene and an inverted repeat sequence of the siRNA. According to the invention, a constructed slow virus vector realizes hereditary silencing to the CXCR4 gene in a T cell; the expression level of the CXCR4 gene in an edited T cell is significantly reduced; and a novel method and thought are provided for treatment of tumors and immune diseases.

An in vitro method for integrating an exogenous DNA sequence into the genome of a cell using a transposon system. The transposon system includes a first vector carrying inverted repeat sequences and an exogenous DNA, and a second vector that expresses a transposases. Also provided is a kit including the transposon system for integrating an exogenous DNA into the genome of a cell. A method for treating a subject is described that includes engineering an immune cell to carry an exogenous DNA sequence using the method and / or the kit described above and administering an effective amount of the engineered immune cell to the subject.

The invention discloses a DNA fragment for inhibiting the expression of an omega secaline gene in a wheat 1B / 1R translocation line and application thereof. The DNA fragment consists of an omega secaline gene coding region partial sequence, a plant intron sequence and the inverted repeat sequence of the omega secaline gene coding region partial sequence which are connected in turn. An RNAi geneticexpression vector containing the DNA fragment is transferred to the wheat 1B / 1R translocation line in a transgenic way, so that the expression of the endogenous omega secaline gene can be inhibited, the adverse effects of omega secaline on the food processing quality of the wheat 1B / 1R translocation line are relieved or eliminated and the food processing quality of the wheat 1B / 1R translocation line is improved.

A novel plant transposon that induces bud mutation, transposase thereof, and a method for efficiently inducing bud mutation by improvement in the transposition efficiency are provided. A novel transposon having terminal inverted repeat sequences (SEQ ID NO: 1 and SEQ ID NO: 2) shown below has been isolated for the first time from carnation (Dianthus caryophyllus) and the nucleotide sequence thereof has been determined. The transposon has contained a gene encoding the transposase in its sequence. The transposon induces bud mutation when it is inserted into a gene. The transposition frequency is elevated due to environmental stress. TAGAGCTGGCAAA—TTTGCCAGCTCTA (SEQ ID NOS 1 & 2, respectively in order of appearance).

The present invention relates to an artificial non-coding RNA containing an inverted SINEB2 repeat sequence and its use in enhancing the translation of a target protein. Specifically, the present invention provides a nucleic acid molecule, which is: (1) an RNA molecule consisting of the following elements: a pairing region complementary to the target mRNA, an inverted SINEB2 sequence, and an optional poly(A) signal region , and optionally regulatory elements; or (2) a DNA molecule capable of transcribing the RNA molecule.