43 results about "UniGene" patented technology

The invention discloses a rhododendron fortunei SSR primer pair based on transcriptome sequencing development, a screening method and application, and belongs to the technical field of biology. The primers are obtained on the basis of transcriptome sequencing and sequence analysis, on the basis of transcriptome sequencing, a large number of data are screened, a unigene sequence which is rich in SSR sites is acquired and SSR labeled primers are designed, and the barriers of a small number of rhododendron fortunei SSR molecular markers and low development efficiency at present are overcome. By arhododendron fortunei group, the effectiveness of the SSR primers is verified, and a foundation is laid for genetic research such as rhododendron fortunei genetic diversity research, genetic linkagemap, QTL positioning and molecular marker-assisted breeding.

The invention provides a method for developing SSR primers of Blumea balsamifera based on transcriptome sequencing. The method comprises: obtaining a Blumea balsamifera genome transcription set, and forming a sequence database; splicing the sequenced sequences by using Trinity to form a transcriptome, and taking the longest transcripts in each gene as Unigene; carrying out Unigene sequence bioinformatics analysis; carrying out SSR detection on the Unigene by using MISA1.0; and carrying out SSR primer design by using Primer3, and carrying out SSR primer polymorphism identification. According tothe present invention, with the method, 17979 pairs of the SSR primers are successfully designed, the 30 pairs of the primers related to the metabolic pathway of the active ingredient are screened, the DNA of 9 Blumea balsamifera with different sources are verified, 27 pairs of the polymorphic primers are determined, and the plant materials Blumea balsamifera with different geographical sources can be distinguished with the 27 pairs of the SSR primers; and the method provides the new idea for the development of the SSR primers of Blumea balsamifera.

The present invention relates to an antifungal protein gene and cDNA sequence thereof, which is obtained by mining the whole genome sequences of Monascus pilosus BCRC 38072 and the unigene database. The gene can encode an antifungal protein MAFP1. A purified protein obtained from M. pilosus culture broth having molecular weight of about 7 kDa is identified as MAFP1 by N-terminal protein sequencing and comparative analysis. The purified MAFP1 protein can inhibit the growth of pathogens such as Paecilomyces variotii BCRC 33174 and Helminthosporium panici BCRC 35004. In addition, it is found by PCR test that the gene of this antifungal protein exists in other Monascus species such as M. Barkeri, M. floridanus, M. lunisporas, M. pilosus, M. ruber and the like. It is also been proved that the mafp1 gene and cDNA thereof in four Monascus strains, M. pilosus (BCRC 38072, BCRC 38093 and BCRC 31502) and M. ruber BCRC 31533, have the same DNA sequences.

The present invention relates to an antifungal protein gene and cDNA sequence thereof, which is obtained by mining the whole genome sequences of Monascus pilosus BCRC 38072 and the unigene database. The gene can encode an antifungal protein MAFP1. A purified protein obtained from M. pilosus culture broth having molecular weight of about 7 kDa is identified as MAFP1 by N-terminal protein sequencing and comparative analysis. The purified MAFP1 protein can inhibit the growth of pathogens such as Paecilomyces variotii BCRC 33174 and Helminthosporium panici BCRC 35004. In addition, it is found by PCR test that the gene of this antifungal protein exists in other Monascus species such as M. Barkeri, M. floridanus, M. lunisporas, M. pilosus, M. ruber and the like. It is also been proved that the mafp1 gene and cDNA thereof in four Monascus strains, M. pilosus (BCRC 38072, BCRC 38093 and BCRC 31502) and M. ruber BCRC 31533, have the same DNA sequences.

The invention provides an SSR marker for detecting a bruchid-resistant variety of broad beans and application of the SSR marker. According to the invention, a PacBio three-generation full-length sequencing technology is combined with an RNA-seq method to carry out whole genome sequencing onbroad bean varieties, software MISA is combined to search Unigene of a transcriptome to obtain a specific broad bean genome sequence containing an SSR core motif, primers are designed according to the SSR core motif, and are used for amplifying an SSR marker, the nucleotide sequence of the upstream primer is shown as SEQ ID NO.1, and the nucleotide sequence of the downstream primer is as shown in SEQ ID NO. 2; the SSR marker primer pair is used for amplifying a broad bean genome template, a PCR amplification product is subjected to polyacrylamide gel electrophoresis, the banding pattern of the PCR product is detected, and the broad bean material for identifying and detecting is the bruchid-resistant material; and the method for detecting the bruchid resistance of broad beans is reliable, simple, convenient and practical, and has an important application prospect in broad bean germplasm resource identification and molecular marker-assisted breeding selection.

The invention provides a method for obtaining transcriptome and functional genes of blumea balsamifera. The method comprises (a) extracting the total RNA(ribonucleic acid) of blumea balsamifera, separating out mRNA (messenger RNA) with polyA (polyadenylation) at the 3' end, randomly breaking the mRNA, performing inverse transcription to synthesize double-stranded cDNA (complementary DNA); (b) sequencing an obtained sequence; (c) splicing and assembling a sequencing result to obtain Unigene and determining the orientation of the Unigene; (d) performing bioinformatics analysis on an obtained gene transcript to obtain the transcriptome and the function genes of blumea balsamifera. The method for obtaining the transcriptome and the functional genes of blumea balsamifera can help obtain 48197273 pieces of sequence information and 100341 Unigenes, involving 60477 pieces of information including RNA-seq names, sequence length and expression quantity, COG (cluster of ortholog genes) predication, COG functional annotations, KEGG (Kyoto encyclopedia of genes and genomes) annotations, KEGG-pathway and GO (gene ontology) annotations, and 37283 pieces of information protein function annotationsfor performing CDS (coding sequence) prediction on the obtained sequence information.

The present invention provides a method for developing elderberry SSR primers based on transcriptome sequencing, the method comprising: S1. Constructing an elderberry transcriptome library and sequencing, splicing and assembling the sequencing results into a complete transcriptome, taking each The longest transcript in the gene is used as Unigene, and the mixed Unigenes library is obtained by splicing; S2. Perform SSR detection on the mixed Unigenes library in step S1; S3. Design SSR primers and identify polymorphisms of the SSR primers. A total of 64,148 pairs of SSR-specific primers were designed in the present invention, and the optimal system of western elderberry fluorescent SSR-PCR was obtained through the experimental optimization scheme. Among the 100 pairs of fluorescently labeled SSR primers synthesized, 25 pairs of primers had good polymorphism. The method for developing elderberry SSR primers based on transcriptome sequencing in the present invention is fast and accurate.

The invention discloses a screening method and application of the SSR molecular marker of the target gene of Acanthopanax. The specific steps of the screening method are: 1) extracting the total RNA of each tissue of Acanthopanax anticana; 2) performing RNA- Seq sequencing, Trinity software for unigene assembly; 3) Screen out unigenes related to the synthesis and accumulation of eleutherosides, SOD, etc., and MicroSatellite software screens the SSR sites on these unigenes; 4) Primer3.0 software designs primers and synthesizes them; 5) Genomic DNA is used as a template for PCR amplification; 6) PCR products are detected by electrophoresis, and SSR primers for the target gene are developed. The invention efficiently and more targetedly develops candidate SSR molecular markers related to target traits, and provides effective markers for molecular identification of excellent Acanthopanax germplasm resources. It is of great significance to the early identification of excellent Acanthopanax germplasm resources and to speed up the breeding process.

The invention discloses a pair of SSR primers for Rhododendron versicolor developed based on transcriptome sequencing and a screening method and application. Belongs to the field of biotechnology. The primers are obtained based on transcriptome sequencing and sequence analysis. On the basis of transcriptome sequencing, a large amount of data is screened to obtain unigene sequences rich in SSR sites and design primers for SSR markers, which overcomes the current SSR molecular The obstacles of small number of markers and low development efficiency. The validity of the SSR primers was verified by Rhododendron versicolor population, which laid a foundation for the genetic diversity research, genetic linkage map structure, QTL mapping, molecular marker-assisted breeding and other genetic research of Rhododendron versicolor.

The invention belongs to the field of molecular biology DNA (Deoxyribonucleic Acid) marker technology and application, and particularly relates to specific primers and a screening method for an EST-SSR (Express Sequence Tag-Simple Sequence Repeat) marker of a torreya grandis transcriptome sequence. The screening method comprises the following steps: firstly, assembling fine torreya grandis seed kernel and circular torreya grandis seed kernel transcriptome data by adopting Trinity software to obtain a transcript sequence; secondly, carrying out SSR site research on Unigene with 1kb or above obtained by screening by using an MISA (MIcroSAtellite identification tool) program, and carrying out screening and separating on 2 to 6 base repeat units and repeat times according to microsatellite fragments with repeat times of 9, 8, 5, 5 and 5 in sequence, thereby obtaining an ESTs sequence with microsatellite repeat; thirdly, designing the primers by using a Primer 3 program; finally, comprehensively evaluating repeatability, stability and polymorphism of the EST-SSR marker to obtain the EST-SSR marker. The specific primers disclosed by the invention can be used for conveniently and quickly analyzing germ plasm resources of different types and genetic diversity of interspecific and intraspecific torreya grandis.

The invention belongs to the technical field of molecular biology, and specifically relates to a PtMBL gene of Portunus trituberculatus mannose-binding lectin, its encoded protein and its application. The present invention uses the unigene and RACE technology obtained by transcriptome sequencing to amplify the PtMBL gene cDNA from Portunus trituberculatus, and finds that the recombinant PtMBL protein has significant bacteriostasis, bacterium binding and bacterium agglutination activities. The recombinant protein PtMBL has obvious inhibitory effects on Gram-negative bacteria (Vibrio alginolyticus, Pseudomonas aeruginosa) and Gram-positive bacteria (Staphylococcus aureus, Micrococcus luteus), and the minimum inhibitory concentrations are 0.81~1.61μM, 0.81μM, 0.81~1.61μM, 3.22~6.44μM. The recombinant protein PtMBL has obvious binding activity to Vibrio alginolyticus, Pseudomonas aeruginosa, Staphylococcus aureus, Micrococcus luteus and Pichia pastoris. In addition, in Ca 2+ In the presence of the recombinant protein PtMBL, it has obvious agglutination effect on Vibrio alginolyticus, Pseudomonas aeruginosa, Staphylococcus aureus, Micrococcus luteus and Pichia pastoris. The invention lays a foundation for the disease prevention and treatment of portunus trituberculatus, and has potential application value in the development of antibacterial drugs, bacterial agglutination preparations, new immune preparations, feed additive production and the like.

The invention discloses a pair of SSR primers developed based on transcriptome sequencing and a screening method and application. Belongs to the field of biotechnology. The primers are obtained based on transcriptome sequencing and sequence analysis. On the basis of transcriptome sequencing, a large amount of data is screened to obtain unigene sequences rich in SSR sites and design primers for EST‑SSR markers, which overcomes the current problems The low number and low development efficiency of red SSR molecular markers are obstacles. The validity of the EST-SSR primers was verified by the Mangospermum population, which laid a foundation for the genetic diversity research, genetic linkage map structure, QTL mapping, molecular marker-assisted breeding and other genetic studies of Mangosteen.

The invention discloses a construction method of Plectropomus microsatellite DNA molecular markers. According to the method, Unigene containing Plectropomus microsatellite repeat sequences is firstly screened out by transcriptome sequencing. Then microsatellite sites are detected in the Unigene sequences. Specific primers of microsatellite markers are designed according to the sequences of both ends of the microsatellite sites and are used for detecting polymorphism of the microsatellite sites. The microsatellite markers with abundant polymorphism of Plectropomus are exploited, and seven microsatellite DNA molecular markers of Plectropomus are verified further. The Plectropomus microsatellite DNA molecular markers provided by the invention can be applied to population genetic structure and genetic breeding study of Plectropomus population.