44 results about "UniGene" patented technology

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 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.