31 results about "Cloning genes" patented technology

The present application relates to the technical field of tomato genetic breeding, and more specifically to molecular markers associated with tomato traits under low-phosphorus stress and their applications. The molecular marker comprises a nucleotide sequence formed by a single nucleotide A>G mutation at position 52526904 of chromosome 11 of the tomato genome. Using this molecular marker, not only can the plant height, fresh weight, and dry weight of tomatoes at the seedling stage under low-phosphorus stress be predicted and identified, but the degree of low-phosphorus tolerance of the plant can also be determined. This molecular marker can assist in cloning genes controlling plant height, fresh weight, and dry weight of tomatoes at the seedling stage under low-phosphorus conditions and in breeding tomatoes for low-phosphorus tolerance, and has important application value.

This application belongs to the field of plant molecular biology technology and specifically provides a PsbHLH35 gene and encoded protein, and their use in tree peony somatic embryogenesis. A PsbHLH35 gene, comprising the nucleotide sequence shown in SEQ ID NO. 1. A protein encoded by the PsbHLH35 gene, comprising the amino acid sequence shown in SEQ ID NO. 2. A method for using the PsbHLH35 gene in tree peony somatic embryogenesis, comprising the following steps: S1) cloning the PsbHLH35 gene into a vector to construct a PsbHLH35 gene overexpression vector; S2) transforming the PsbHLH35 gene overexpression vector into tree peony. The PsbHLH35 gene of this application has a significant promoting effect on tree peony somatic embryogenesis.

The invention relates to a brassica napus drought-resistant cultivation method based on a target gene, which comprises the following steps: S1, cloning a BnaWRKY038 gene, and the nucleotide sequence of the BnaWRKY038 gene is as shown in SEQ ID NO: 1; s2, constructing an overexpression vector of the BnaWRKY038 gene, and transforming agrobacterium tumefaciens; s3, performing mediation transformation on the hypocotyl of the brassica napus by utilizing the agrobacterium to obtain a transgenic plant; s4, screening and identifying the transgenic plants to obtain the drought-resistant brassica napus with the overexpressed BnaWRKY038 gene. By obtaining the BnaWRKY038 overexpressed transgenic plants and based on phenotype results of the overexpressed transgenic rape plants, the drought resistance of the plants can be positively regulated under drought treatment of the rapes, a new choice is provided for screening dominant stress-resistant genes, and the drought-resistant brassica napus with the overexpressed BnaWRKY038 gene is obtained. The method has important application value in the field of plant genetic engineering, can effectively reduce economic loss caused by drought, and promotes cultivation of new varieties of drought-resistant rape.

The invention provides an OsSPL17 gene for improving the strength of rice stalks and application of the OsSPL17 gene, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the OsSPL17 gene disclosed by the invention is as shown in SEQ ID NO. 1. According to the invention, a transgenic vector is constructed by cloning a promoter and a genome segment of an OsSPL17 gene, and transgenic rice of which an OsSPL17 self-promoter drives an OsSPL17 self-genome is obtained. Experimental results show that compared with a wild type, the stem thickness, the stem wall thickness and the stem breaking resistance of the transgenic rice are remarkably increased, which indicates that the OsSPL17 plays an important regulation role in rice stem development and can be used for cultivating lodging-resistant rice varieties.

This application relates to the technical field of tomato genetic breeding, and specifically to molecular markers associated with low-temperature tolerance in tomato seedlings and their applications. This molecular marker comprises an Indel deletion sequence from positions 6494531 to 6494551 on chromosome 5 of the tomato genome, SL2.50ITAG2.4. This molecular marker enables accurate detection of low-temperature tolerance in tomato seedlings. Furthermore, this application provides closely associated molecular markers for cloning genes that control the degree of leaf wilting caused by water loss in tomato seedlings under low-temperature stress, and for breeding tomato varieties for low-temperature tolerance, thus possessing significant application value.

This invention discloses a photoperiod regulation gene of foxtail grass. SvEhd1 Its application in shortening the research cycle of rice resistance to rice blast falls under the fields of genetic engineering and plant breeding. SvEhd1 The gene nucleotide sequence is shown in SEQ ID NO.1. This invention utilizes cloning... SvEhd1 Genes and their promoters were used to construct recombinant expression vectors, which were then transformed into the blast-susceptible rice variety "Taipei 309," resulting in transgenic rice materials with significantly shortened growth periods. Under natural and artificial long-day conditions, the heading time of the transgenic plants was significantly earlier than that of the wild type, the generation rate was accelerated, and the blast susceptibility of the original variety was not altered. The transgenic materials and methods provided by this invention can effectively shorten the experimental cycle for verifying the function of rice blast resistance genes and evaluating resistance, providing an efficient experimental system for rice disease resistance research.

The present application relates to the field of plant growth and development molecular biology, and particularly relates to a gene OsARF14b and application thereof in controlling rice root elongation and tillering, the gene OsARF14b has the nucleotide sequence shown in SEQ ID NO:1, and the application of the gene OsARF14b in regulating rice root elongation and tillering traits.The present application obtains overexpression and knockout transgenic materials by cloning the gene OsARF14b, constructing a transgenic vector, and statistically analyzing the phenotype of the transgenic materials, and finds that the root length and tiller number in different materials change significantly, compared with the wild type (Zhonghua 11, ZH11), the root length of the knockout transgenic material (cas9) is significantly shortened, and the tiller number is significantly increased, the root length of the overexpression material (OE) is lengthened, and the tiller number is significantly reduced, which indicates that the OsARF14b gene plays an important regulatory function in the process of plant root elongation and tillering.

The invention provides a tobacco chloroplast polypeptide Ntppc and application thereof, a tobacco chloroplast related polypeptide named Ntppc is separated and identified, an overexpression vector is constructed and tobacco is transformed by cloning an Ntppc gene (SEQ ID NO: 2), and experiments prove that the susceptibility of tobacco to black shank is remarkably enhanced by overexpression of the gene. The invention provides a new mechanism for plant immune regulation and control, and has important agricultural application value.

The application discloses a PtrSAUR32 gene and application thereof in enhancing growth of citrus root, relates to the field of agricultural biological genes, and solves the technical problem of a technical blank in the prior art in regulating growth of the citrus root by using SAUR; the coding sequence of the PtrSAUR32 gene is a nucleotide sequence as shown in SEQ ID NO.1; the transgenic plant with enhanced root growth is obtained by cloning the coding sequence of the PtrSAUR32 gene, constructing a PtrSAUR32 overexpression vector, genetically transforming the citrus by using the PtrSAUR32 overexpression vector, and identification; the application improves the expression level of the PtrSAUR32 based on the PtrSAUR32 overexpression, and obtains the transgenic citrus plant with enhanced root growth; the root length of the plant is significantly higher than that of a control plant, and the plant has great application value in improving the citrus rootstock and enhancing the stress resistance of the citrus.

This invention discloses the promoter of the rice Os04g0650800 gene and its applications, belonging to the field of plant biotechnology. By cloning the promoter sequence of the Os04g0650800 gene, a recombinant expression vector pPGDH-YFP driving the yellow fluorescent protein gene was successfully constructed. Positive rice plants were screened by infecting Nipponbare rice with Agrobacterium. The T0 generation of positive rice seedlings were inoculated with Xanthomonas oryzae pv. oryzae. Xoo Using a confocal microscope, it was discovered that... Xoo Infection of positive rice plants can induce YFP expression. Genetic engineering of rice using this promoter, such as regulating the efficient expression of target genes through this promoter, will yield positive results. Xoo Infected rice plants; if this promoter is used to drive the expression of disease resistance-related genes in rice, rice plants resistant to bacterial blight will be obtained.

This invention relates to the cloning and application of the salt tolerance gene LpWRKY75 in Lilium pumilum DC. The steps include: (1) cloning of the target gene; (2) construction of the expression vector; and (3) detection of physiological indicators in transgenic plants. This experiment used Lilium pumilum as the research object, cloning the LpWRKY75 gene from the cDNA of Lilium pumilum roots using homologous recombination technology; transforming tobacco by constructing an overexpression vector; and identifying the function of LpWRKY75 overexpression in improving tobacco salt stress through transgenic tobacco. The function of the LpWRKY75 gene in plant salt stress tolerance was preliminarily clarified, laying the foundation for further application of Lilium pumilum in lily breeding and providing important candidate gene resources for plant stress resistance breeding.

The invention provides an SD5 gene for regulating and controlling the plant height and yield of rice and application of the SD5 gene, and relates to the technical field of plant genetic engineering. The nucleotide sequence of the whole genome of the SD5 gene is as shown in SEQ ID NO. 1, and the nucleotide sequence of the coding region of the SD5 gene is as shown in SEQ ID NO. 2. A transgenic vector is constructed by cloning a promoter sequence of a DEP1 gene and a coding region sequence of an SD5 gene, and transgenic rice of which the SD5 gene is driven by a promoter of the DEP1 gene is obtained. Experimental results show that compared with a transformation receptor ZH17, the plant height of a transgenic line is reduced by 8.1%, the number of grains per spike of a main stem is increased by 9.6%, the thousand seed weight is increased by 4.3%, and the yield of a single plant is increased by 15.9%. Therefore, the SD5 gene plays an important role in regulating and controlling the plant height and the yield of the rice and can be used for cultivating lodging-resistant rice varieties and high-yield rice varieties.

The invention discloses application of a transcription factor GhBBX21 in promoting anthocyanin accumulation in R1 cotton, and belongs to the technical field of biology. A gene CDS sequence is cloned, an overexpression vector 35S: GhBBX21-GFP is constructed, wild type arabidopsis (WT) is transformed by a pseudomonas inflorescence infection method, and an overexpression plant is obtained. Yeast single hybrid experiments prove that GhBBX21 can be combined on a promoter of an important transcriptional gene GhPAP1D for anthocyanin synthesis of R1 cotton. A dual luciferase experiment proves that the GhBBX21 can be used for remarkably promoting the expression of GhPAP1D in the R1 cotton. The results provide effective support for mining important genes related to cotton anthocyanin synthesis and analyzing a cotton anthocyanin synthesis route, and provide important support for improving cotton resistance and increasing cotton ornamental value by utilizing the genes, creating excellent cotton planting resources and cultivating insect-resistant cotton and ornamental cotton varieties.

The application discloses a plant-derived plastic-degrading enzyme and a method for degrading PP plastic in vitro, and belongs to the technical field of enzyme engineering. The plant-derived plastic-degrading enzyme is a plant-derived HIS1 protease. The method for degrading PP plastic in vitro comprises the following steps: cloning a HIS1 gene from a plant by using an RT-PCR technology; connecting the gene to a prokaryotic expression plasmid by using a blunt-end cloning technology; introducing the gene into an expression platform of Bl21 (DE3) E. coli to express and purify the HIS1 protein; and degrading PP plastic in an enzyme reaction environment constructed in vitro. The HIS1 protein provided by the application can effectively degrade PP plastic, provides a new idea for degrading plastic by using plant enzymes, and has a good application prospect.

The application discloses a genetic element for enhancing lactic acid metabolic flux of actinomycetes, and belongs to the field of synthetic biology and microbial metabolic engineering. sucC The application successfully constructs a single expression element and a co-expression element by cloning genes from Streptomyces coelicolor sucD , and verifies that the SucC protein and the SucD protein both have lactic acid coenzyme A synthetase activity through in-vitro protein purification and enzyme activity determination, and the enzyme activity of a protein complex (SucC-SucD, SucCD) expressed by the co-expression element is significantly higher than that of the single protein, indicating that the two proteins have a synergistic effect; the genetic element can efficiently catalyze the combination reaction of lactic acid and coenzyme A in vitro, accelerates the lactic acid metabolic speed, provides a core genetic element for the modification of the lactic acid metabolic pathway of actinomycetes, enriches a synthetic biology tool library, and has important industrial application prospects and academic value.

The application discloses a method for creating cotton compact plant type material by using a gene editing technology, that is, cloning a GhGRF2 gene from Gossypium hirsutum, designing a gene editing site, constructing a gene editing vector, performing genetic transformation by using a cotton agrobacterium mediation method, obtaining a transgenic regenerated plant, performing positive identification on the transgenic plant, analyzing mutation of the editing site, observing a phenotype of the gene editing mutant plant, and further obtaining compact germplasm material which can be used for cotton plant type breeding. The application belongs to the technical field of plant biotechnology, and specifically relates to a method for creating cotton compact plant type material by using a gene editing technology, wherein a cotton GhGRF2 gene is knocked out by using a CRISPR-Cas9 gene editing technology, and cotton compact plant type material is obtained, so that the problem that it is difficult to improve cotton plant type by using a traditional breeding method is solved.

This invention belongs to the field of genetic engineering technology, specifically relating to a BAMT gene that regulates salt-alkali tolerance in sunflowers and its application. The BAMT gene has the nucleotide sequence shown in SEQ ID NO.1. The amino acid sequence of the expressed protein of the BAMT gene is shown in SEQ ID NO.2. The specific primer pair nucleic acid sequences required for cloning the BAMT gene are shown in SEQ ID NO.3 and SEQ ID NO.4, or in SEQ ID NO.5 and SEQ ID NO.6. This invention also discloses the application of the BAMT gene regulating salt-alkali tolerance in sunflowers in a kit. To verify the difference in salt-alkali tolerance between two sunflower hybrids, hydroponic seedlings were treated with a saline-alkali solution. The results showed that the BAMT gene was continuously upregulated in K-27 and continuously downregulated in K-7, exhibiting a significant variety-specific expression pattern. This reveals the physiological and transcriptional regulatory characteristics of sunflowers in response to combined salt and alkali stress, and the identified BAMT gene can provide a reference for molecular breeding for salt-alkali tolerance.

A system for the molecular detection and genotyping of Toxoplasma gondii, consisting of: a DNA extraction unit configured to extract and purify DNA from stool and tissue samples; a PCR amplification unit configured to amplify T. gondii -specific DNA sequences using the nested PCR technique with primer sets comprising: external primers sag2Fext and sag2Rext, which target the non-coding region of the SAG2 gene to generate 1000 bp amplicon and nested inner primers sag2Fint and sag2Rint targeting the coding region of the SAG2 gene to generate a 500 bp amplicon; a gel electrophoresis system configured to visualize PCR amplification products on agarose gel; a restriction digestion unit configured to digest the nested PCR products with BsrDI and BspMI enzymes in a single reaction for genotyping; and a genotype analysis unit configured to classify the genotype of T. gondii into three clonal lineages and non-clonal genotypes based on restriction fragment profiles.

The invention discloses an application of a soybean PSK oligopeptide gene GmPSK alpha 14 in regulation and control of soybean growth and phosphorus nutrition thereof. The invention provides new application of the GmPSK alpha14 gene, and research shows that the GmPSK alpha14 gene can respond to low-phosphorus stress, is obviously subjected to up-regulation expression of low-phosphorus stress in soybean roots and leaves, and participates in regulation and control of soybean phosphorus nutrition and growth under low-phosphorus stress; the GmPSK alpha14 gene is cloned to prepare a recombinant vector, the recombinant vector is transformed into soybean for overexpression, it is shown that the GmPSK alpha14 gene is capable of regulating plant phosphorus nutrition, under the condition of phosphorus nutrient deficiency, overexpression of the GmPSK alpha14 gene can improve the phosphorus deficiency symptom of soybean, growth of soybean and soybean root systems can be remarkably promoted, absorption of soybean to phosphorus nutrient and accumulation of biomass can be improved, and the yield of the soybean can be increased. The plant phosphorus content is increased, the soybean growth is regulated, and more gene resources are provided for the plants to adapt to environmental low-phosphorus stress.

The present application relates to the field of plant growth and development molecular biology, and particularly relates to genetic engineering application of gene OsDLN194, the gene OsDLN194 has the nucleotide sequence shown in SEQ ID NO.1, the protein sequence of the gene is SEQ ID NO.2, and the gene is located on the 7th chromosome of rice.The application of gene OsDLN194 in regulating at least one trait of rice tillering, yield per plant and nitrogen absorption.The present application obtains overexpression plants by cloning gene OsDLN194, constructing a transgenic vector, determining the phenotype of the transgenic plants, finds that the tiller number of the overexpression plants is significantly changed, and compared with wild type rice, the overexpression plants show the traits of increased tiller number, increased yield per plant, improved nitrogen absorption capacity and the like, which indicates that OsDLN194 gene plays an important regulatory function in the process of rice tillering, yield per plant and nitrogen absorption.

The present application relates to the field of biotechnology, in particular to a Millettia pachynema gene LpDUF707-1 and application thereof, and is based on Millettia pachynema genome data and BSA resequencing data, cloning of the LpDUF707-1 gene, and analysis of the expression pattern under drought stress by using real-time quantitative PCR technology, and it is found that the LpDUF707-1 gene can promote plant root growth and improve the drought tolerance of plants, and the present application lays a foundation for revealing the drought tolerance mechanism of the LpDUF707-1 gene, and provides gene resources and theoretical basis for plant drought tolerance molecular breeding.

The present application relates to the field of biotechnology, in particular to a white flower carum gene MaUGT79 and its application, the present application is based on white flower carum genome data and BSA resequencing data, cloning MaUGT79 gene, using real-time quantitative PCR technology to analyze the expression pattern under drought stress, and construct the recombinant prokaryotic expression vector to verify MaUGT79 gene function, it is found that transgenic escherichia coli can catalyze scopolin to produce scopolamine; construct plant overexpression vector and RNAi vector to transform the gene into white flower carum hairy root, verify MaUGT79 gene to participate in scopolin synthesis, has the function of resisting drought stress, the present application lays the foundation for revealing the drought tolerance mechanism of MaUGT79 gene, provides gene resources and theoretical basis for plant drought tolerance molecular breeding.

The invention relates to clone of a salt-tolerant gene LpWRKY75 of Lilium pumilum DC, and an application of the salt-tolerant gene LpWRKY75 of Lilium pumilum DC. Comprising the following steps: (1) cloning a target gene; (2) constructing an expression vector; and (3) detecting physiological indexes of the transgenic plant. According to the experiment, lilium tenuifolium is taken as a research object, and the LpWRKY75 gene is cloned from cDNA (complementary deoxyribonucleic acid) at the root of the lilium tenuifolium by using a homologous recombination technology; transforming tobacco by constructing an overexpression vector; through transgenic tobacco identification, overexpression of the LpWRKY75 is used for improving the salt stress function of the tobacco, the function of the LpWRKY75 gene in salt stress resistance of the plant is preliminarily clarified, a foundation is laid for further application of lilium tenuifolium in lily breeding, and an important candidate gene resource is provided for stress resistance breeding of the plant.

The invention discloses a non-cloning genomics analysis method based on a SCoT marker, which can directly restore 01 data into a simulation sequence of PCR (Polymerase Chain Reaction) according to an electrophoretogram by adopting a new data processing and statistical analysis method'primer replacement method 'without cloning and sequencing after completing SCoT electrophoresis on a to-be-detected target. Homologous genes are directly retrieved through bioinformatics analysis of SCoT sequences, classification analysis and function analysis are carried out according to the copy number, existence and distribution of SCoT differential genes, and the molecular basis of genetic diversity variation of genomes is completely revealed. According to the method, SCoT marker analysis is improved into SCoT omics analysis, bioinformatics analysis can be carried out by utilizing genomics big data, a molecular basis of genomic genetic diversity variation among different materials is disclosed, and the method has important significance and wide application prospects in improving the breeding efficiency, accelerating the breeding process and improving the fundamental research theory level.

The application is a divisional application of 202011444476.6. The application relates to the field of plant growth and development molecular biology, in particular to the application of gene OsPIN10b in plant root elongation, the gene OsPIN10b has a nucleotide sequence shown in SEQ ID NO:1, and the application of the gene OsPIN10b in regulating the length of plant roots. By cloning the gene OsPIN10b, a transgenic vector is constructed, overexpression and knockout transgenic materials are obtained, the phenotype of the transgenic materials is measured, the root morphogenesis of different materials is significantly changed, the overexpression materials have the characteristics of increased root length and the like compared with the knockout transgenic materials and wild type materials, and it is indicated that the OsPIN10b gene plays an important regulating function in the development process of rice root elongation.

The application discloses application and a method of a radish RsBBX9 gene in regulation of anthocyanin synthesis, and belongs to the technical field of plant genetic engineering. The application verifies the regulation of RsBBX9 on anthocyanin content by cloning the RsBBX9 gene, constructing an overexpression vector pFGC1008-RsBBX9 and a VIGS silencing vector pTY-S-RsBBX9, combining transient expression and gene silencing technology. The application further verifies the interaction of RsBBX9 and RsMYB1 by a yeast double-hybrid experiment, a BiFC experiment and a pull-down experiment, and the interaction cooperatively activates an anthocyanin synthesis path. The application provides an efficient technical scheme for radish variety improvement and anthocyanin production.

The invention discloses a cultivation method of drought-resistant brassica napus, and belongs to the technical field of biology. The method comprises the following steps: by taking brassica napus as a research object, extracting leaf RNA (Ribonucleic Acid), performing reverse transcription to obtain cDNA (Complementary Deoxyribonucleic Acid), cloning a BnaNCED3 gene, constructing an overexpression and silence vector, performing transformation verification by escherichia coli, and introducing agrobacterium; infecting the oilseed rape hypocotyl explant through agrobacterium tumefaciens, and carrying out co-culture, differentiation screening and rooting culture to obtain a transgenic plant; and finally, screening strains with high expression of BnaNCED3, and identifying the drought tolerance. By improving the expression level of the BnaNCED3 gene, the drought survival rate of the oilseed rape is remarkably improved, the water loss rate of leaves is reduced, and an efficient technical means is provided for cultivation of drought-resistant oilseed rape varieties.

This invention discloses a barley stripe disease resistance gene. Rdg2b This invention relates to the field of bioengineering technology and its applications. It describes the cloning of [specific species / organisms] from Ganbei No. 2 barley. Rdg2b The complete CDS segment of the gene was extracted, and an overexpression vector was constructed. Agrobacterium-mediated transformation was then performed to obtain overexpression-positive lines. Infection experiments with *Strombus striatum* showed that, compared to the wild type, [the expression was positive]. Rdg2b Overexpression lines showed significantly enhanced disease resistance. Real-time quantitative PCR analysis revealed that overexpression lines exhibited... Rdg2b Gene expression was significantly upregulated, confirming Rdg2b This invention positively regulates barley disease resistance. It provides important genetic resources for molecular breeding of barley for disease resistance.

The application discloses a fruit color regulation gene MdMYBS1 and application thereof, relates to the technical field of genetic engineering, and has the technical scheme as follows: a fruit color regulation gene MdMYBS1, the nucleotide sequence of the MdMYBS1 gene is shown in SEQ ID NO.1, and the amino acid sequence is shown in SEQ ID NO.2; after overexpression of the MdMYBS1 gene in apple callus, fruit, tissue culture seedlings and tomatoes, yellow color is deepened, the content of carotenoids is significantly increased, and the fruit color can be regulated; by cloning the gene MdMYBS1, the overexpression vector and the interference vector of MdMYBS1 are constructed, genetic transformation is carried out on apple fruit, callus or tissue culture seedlings as materials, the regulation mode of the gene in fruit carotenoid accumulation is revealed, and directional selection breeding of fruit yellow color regulation is facilitated.

The present invention discloses the overexpression of the CiKPI gene and its application in improving the resistance of citrus to anthracnose, which relates to the field of biotechnology and solves the technical problem that the prior art lacks the research and application of using KPI to improve the resistance of citrus to anthracnose; the encoded protein of the CiKPI gene is a trypsin inhibitor, and the CiKPI gene has the nucleotide sequence shown in SEQ ID NO.1. By cloning the CiKPI gene, constructing a CiKPI overexpression vector, transforming the CiKPI overexpression vector into citrus, and identifying, transgenic citrus plants with improved anthracnose resistance are obtained; through the overexpression of the CiKPI gene, the present invention can significantly reduce the lesion size of fungal anthracnose of citrus and alleviate the incidence degree of citrus anthracnose, and the incidence degree of anthracnose can be reduced to a maximum of 48.8% of the existing citrus.