43 results about "Inverted Repeat Sequences" patented technology

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 relates to a method for cultivating an anti-TYLCV (Tomato Yellow Leaf Curl Virus) tomato plant by using an RNAi (RNA interference) technology, belonging to the field of biotechnology. According to the method, the conserved sequences in genes of V2 and C1 of TYLCV are inosculated, an RNAi carrier with inverted repeat sequences is constructed, the aseptic seedling cotyledons or hypocotyl explants of the tomato are transformed through an agrobacterium-mediated transformation method, the transgenic tomato expresses resistance to the TYLCV after the inoculation of the agrobacterium infectious clone injection of the TYLCV and the TYLCV-carrying Bemisia tabaci virus, and no obvious disease symptoms are observed. According to the method, the anti-TYLCV transgenic tomato plant can be cultivated by using the RNAi technology, so that the technical support for the control of the TYLCV is provided.

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.

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.

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.

A trap vector containing a loxP sequence composed of inverted repeat sequence 1, a spacer sequence and inverted repeat sequence 2 in this order, the loxP sequence being a mutant loxP wherein a part of the inverted repeat sequence 1 or 2 is mutated.

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 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 synthesizedBt 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 relates to the field of biotechnology, and in particular relates to a method for controlling soot lice by silencing two resistance genes.. The method comprises the following steps of: selecting bemisia tabaci drug-resistant CYP6CM1 gene and coe1 gene as RNAi target gene; embedding partial sequences of the CYP6CM1 gene and the coe1 gene by overlap polymerase chain reaction (PCR), and using the embedded sequence to build an RNAi carrier with inverted repeat sequence; and transforming the RNAi carrier into a target plant, and obtaining transgenic plant capable of controlling the bemisia tabaci. The method is an important supplement of a method for controlling the bemisia tabaci mainly by an insecticide. The method can be used for interfering with the expression of related drug-resistant genes in the bemisia tabaci by the RNAi technology, thus providing technical support for controlling the bemisia tabaci.

The present invention provides an adeno-associated virus recombinant vector for knocking out the CXCL12 gene, the recombinant vector at least includes the following sequence elements operably linked: 5' terminal inverted repeat sequence, CMV promoter, sacas9 sequence, polyA signal sequence, U6 promoter sequence, gRNA sequence, 3'-end inverted repeat sequence. The inventors found that the CXCL12 gene knockout vector can effectively inhibit the growth of transplanted tumors and tumor angiogenesis in the U87 cell glioma model, and can effectively inhibit the tumor growth and tumor angiogenesis in the U87 cell subcutaneous glioma model in nude mice. It shows that the technical scheme of the present invention can be applied to the treatment of glioma diseases.

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.

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.

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.

The invention relates to a fish Tc1-like active transposon and its application. Specifically provided are transposons with complete left and right terminal inverted repeat sequences, transposons capable of encoding transposases and transposases encoded therein, gene vectors and gene transfer systems containing the above transposons and transposases. Through the verification of transposition activity in zebrafish, it is confirmed that the above-mentioned transposon has transposition activity, and the gene transfer system can perform high-efficiency transposition in zebrafish, so the present invention is to further verify the transposition efficiency of transposon And the use of the transposon in vertebrate related research and application has laid a foundation.

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.