673 results about "Enzyme digestion" patented technology

The invention discloses a recombinant humanized XVII type collagen as well as an expression vector and a genetically engineered bacterium thereof. Four functional fragments with high activity are obtained through screening, one of the fragments is optimized and then combined according to a certain sequence to obtain recombinant humanized XVII type collagen JYC1703, or one of the fragments is optimized and then combined according to a certain sequence, and GPP sequences are fused at an amino terminal and a carboxyl terminal respectively to obtain recombinant humanized XVII type collagen JYC1701 and JYC1702, and the recombinant humanized XVII type collagen JYC1701 and JYC1702 are combined according to a certain sequence to obtain the recombinant humanized XVII type collagen JYC1701 and JYC1702. After codon optimization, amplifying and cloning into an expression vector, transforming into pichia pastoris competent cells after enzyme digestion linearization, and screening high-copy recombinants through G418 resistance gradient, so as to obtain pichia pastoris genetically engineered bacteria for expressing recombinant humanized XVII type collagen. The recombinant humanized XVII type collagen JYC1701, JYC1702 and JYC1703 disclosed by the invention show excellent functions of promoting cell adhesion, cell proliferation and cell migration, and are suitable for preparing products such as skin care products or medical instruments and the like.

The invention relates to a cascade gating activated CRISPR / Cas12a system which is used for synchronous detection of miRNA155 and FEN1, the system comprises recognition assembly and enzyme digestion of two DNA hairpins, and Cas12a is activated by an enzyme digestion product to generate a fluorescence signal. The DNA hairpin 1 comprises a recognition part of miRNA155, a hidden foothold of the DNA hairpin 2 and a part of Cas12a activation chain, and the DNA hairpin 2 comprises a part combined with the hidden foothold of the DNA hairpin 1 and a part of Cas12a activation chain. In the application, two DNA hairpins are combined and assembled under the catalysis of miRNA155 to be called as an FEN1 enzyme cutting substrate, a CRISPR / Cas12a system can be activated in a cascade gating manner, and signals are generated only when the two targets exist, so that the specificity synchronous detection of the two targets is completed.

The invention provides a KASP molecular marker related to a corn ear height character under a drought condition and application of the KASP molecular marker, and belongs to the technical field of molecular genetics. The SNP marker site related to the ear height character under the drought condition and having polymorphism is obtained through whole genome association analysis, the SNP marker site refers to a corn B73 RefGenv4 genome, when nucleotide polymorphism T / C exists at the position 147243591 of a 10 # chromosome, the SNP marker site is a dominant character, and nucleotide sequences used for amplifying a KASP molecular marker primer group are shown as SEQ ID NO.1-3. Gene typing identification can be rapidly, simply and efficiently carried out by utilizing a KASP typing technology, enzyme digestion and electrophoresis are not needed, accurate identification and selection can be carried out in a seedling stage, a large amount of breeding workload can be reduced, the period of high-ear-position breeding selection can be shortened, and the method can be used for high-ear-position molecular marker-assisted breeding of corn.

The invention discloses a dCAPS molecular marker for rapidly identifying the content of soybean protein and application of the dCAPS molecular marker. An upstream primer sequence of a primer pair for amplifying the dCAPS molecular marker is as shown in SEQ ID No. 1, and a downstream primer sequence is as shown in SEQ ID No. 2. According to the invention, the mispairing base primer is introduced to carry out PCR amplification and enzyme digestion on the to-be-detected soybean genome DNA, and the soybean protein content can be identified according to the band size of the enzyme digestion product. The dCAPS molecular marker disclosed by the invention can be used for identifying or assisting in identifying the soybean protein content, a high-protein soybean variety and a low-protein soybean variety are distinguished from the molecular level, and the dCAPS molecular marker has important significance on soybean molecular marker assisted breeding and high-protein soybean screening and cultivation.

The invention discloses a method for preparing a simeglutide precursor peptide through intracellular enzyme digestion. The method comprises the following steps: (1) connecting a dissolution-promoting tag GST and the simeglutide precursor peptide together through protease specific recognition of amino acid sites to form a fusion protein; (2) integrating the coding nucleotide sequence of the fusion protein, the coding nucleotide sequence of KEN2 protease and the coding nucleotide sequence of carboxypeptidase B into the same expression vector to form a recombinant vector; (3) transforming the recombinant vector into engineering bacteria and carrying out fermentation production; and (4) centrifuging a fermentation product, collecting thalli, and purifying to obtain the precursor peptide powder of the semeglutide. The whole operation process is simpler, more convenient and smoother, tedious recovery steps are omitted, and the preparation and recovery difficulty of the precursor peptide of the semeglutide is remarkably reduced.

The invention belongs to the technical field of biology, and relates to a recombinant mycobacteriophage capable of enabling host bacteria to express nano luciferase as well as a preparation method and application of the recombinant mycobacteriophage. The recombinant mycobacterium fluorescent bacteriophage capable of enabling the host bacteria to express the nano luciferase comprises an expression element for expressing the nano luciferase in mycobacteria, the expression element contains a gene nluc required by luminescence and a mycobacterium strong promoter, and the upstream of the gene nluc is connected with a promoter sequence to ensure the normal expression of downstream genes. The base sequence is as shown in SEQ ID NO: 1. The preparation method comprises the following steps: carrying out enzyme digestion on a bacteriophage skeleton to form a linear sequence, enabling the linear sequence to be recombined with a target fragment containing a gene nulc, connecting the enzyme-digested bacteriophage skeleton with the target fragment containing the gene nluc, carrying out in-vitro packaging, transferring into an escherichia coli competent cell, and carrying out in-vivo amplification in escherichia coli; the method comprises the following steps: extracting positive phagocytids of recombinant escherichia coli, electrically transferring the phagocytids into Mycobacterium smegmatis, and culturing the phagocytids on a double-layer agar plate to obtain the bacteriophage. According to the application of the recombinant mycobacterium bacteriophage in the mycobacterium rapid detection and / or drug sensitivity detection reagent, the method is simple, convenient and rapid, the sensitivity is high, and the repeatability is strong.

The present application relates to a virus of reverse genetic operation construction of NADC34-like PRRSV2. Specifically, the full gene sequence of NADC34-like PRRSV2 BJ1805-2 isolated strain is segmented and connected to the vector by using PCR amplification, enzyme digestion and other methods with pACY177 plasmid as the carrier, and the BJ1805-2 full-length infectious clone recombinant plasmid (rBJ1805-2) is obtained. The infectious clone virus rBJ1805-2 is rescued in vitro, and the first NADC34-like PRRSV2 strain infectious clone platform in China is successfully built. The present application also relates to a modification method for constructing NADC34-like PRRSV2 strain which can adapt to Marc-145 cell passage culture. The infectious clone virus constructed by the present application has good in vitro and in vivo proliferation efficiency, and can cause specific viremia after inoculation in pigs; it has good safety, does not cause fever after inoculation in pigs, and will not cause piglet death. The modified strain can be used as a candidate strain for developing the first NADC34-like PRRSV2 specific vaccine in China, and is conducive to the prevention and control of PRRSV epidemic in China.

The invention provides a non-disease diagnosis purpose 5-hydroxymethylcytosine 5hmC detection method and a non-disease diagnosis purpose 5-hydroxymethylcytosine 5hmC detection kit. The detection method comprises the following steps: S1, glycosylation treatment; s2, carrying out enzyme digestion treatment on the GSREs; s3, carrying out APOBEC deamination treatment; s4, carrying out hydroxymethylation detection; the kit comprises a glycosylation reaction kit, a GSREs enzyme digestion reaction kit, an APOBEC deamination reaction kit and a PCR (Polymerase Chain Reaction) kit. The glycosylation reaction kit comprises a CutSmart buffer solution, UDP-Glucose, a T4-BGT enzyme and nuclease-free water, wherein the CutSmart buffer solution, the UDP-Glucose and the T4-BGT enzyme are used as raw materials; the GSREs enzyme digestion reaction kit comprises a GSREs enzyme, an enzyme digestion reaction buffer solution and nuclease-free water, the APOBEC deamination reaction kit comprises APOBEC, BSA (Bovine Serum Albumin), an APOBEC Reactive buffer solution and nuclease-free water; the PCR kit comprises a PCR reaction solution, a primer and a probe. According to the 5-hydroxymethylcytosine 5hmC detection method and kit for non-disease diagnosis purpose provided by the invention, precise quantification of extremely high specificity of 5hmC is realized, and the detection sensitivity is also improved while the specificity of a detection signal is improved.

The invention relates to the technical field of plasmid extraction, in particular to an automatic extraction method of endotoxin-free large-volume plasmid DNA and a kit. According to the kit provided by the invention, a magnetic bead adsorption technology and a buffer system with a unique separation effect are adopted, and specially modified protein is combined with magnetic beads, so that endotoxin and protein impurities can be adsorbed; an endotoxin removing solution is adopted, so that endotoxin can be efficiently removed; the method comprises the following steps: firstly, preparing plasmid DNA, then, uniquely embedded nucleic acid adsorption magnetic beads have very strong affinity to the plasmid DNA under a certain condition and are not easy to adsorb endotoxin, and when the condition is changed, the nucleic acid adsorption magnetic beads release the adsorbed plasmid DNA, so that the purpose of rapidly separating and purifying the plasmid DNA is achieved. The plasmid DNA extracted by utilizing the kit and the extraction method disclosed by the invention is high in yield, good in purity and extremely low in endotoxin content, and can be suitable for transfecting various cells and various conventional operations including enzyme digestion, PCR (Polymerase Chain Reaction), sequencing, connection and other various downstream experiment requirements.

The invention provides a quality detection method for mRNA capping rate and PolyA distribution. The quality detection method comprises the following steps: a) mixing mRNA to be detected, a capping rate analysis probe and a PolyA distribution analysis probe to obtain an mRNA-probe structure; b) carrying out enzyme digestion on the mRNA-probe structure by using thermal stable RNase H enzyme to obtain a 5'enzyme digestion fragment and a 3 'enzyme digestion fragment; and c) detecting the 5 '-enzyme digestion fragment and the 3'-enzyme digestion fragment by using liquid chromatography-mass spectrometry, obtaining the capping rate from the 5 '-enzyme digestion fragment, obtaining the PolyA distribution condition from the 3'-enzyme digestion fragment, and realizing simultaneous detection of the 5 '-capping rate and the 3' Poly A. The method can solve the problems of many steps and long time required by detection of the capping rate and the PolyA distribution of mRNA in the prior art, and is suitable for the field of mRNA quality detection.

The invention relates to the technical field of biology, and particularly discloses a fusion protein containing an SUMO tag and a preparation method and application thereof. The fusion protein provided by the invention comprises an SUMO tag and an antibacterial peptide, the antibacterial peptide can inhibit escherichia coli and staphylococcus aureus, and the amino acid sequence of the antibacterial peptide is as shown in SEQ ID No. 10, SEQ ID No. 6, SEQ ID No. 8, SEQ ID No. 12, SEQ ID No. 15 or SEQ ID No. 17. The fusion protein can be efficiently expressed through a prokaryotic expression system, renaturation of the antibacterial peptide can be successfully achieved after enzyme digestion and purification, the antibacterial activity of the antibacterial peptide is recovered, and therefore efficient preparation of the antibacterial peptide is achieved.

The invention provides a high-throughput single-cell multi-omics three-dimensional genome detection method and system. The detection method comprises the following steps: enzyme digestion reaction of fixed cells; carrying out proximity ligation treatment on the cells subjected to the enzyme digestion reaction; carrying out analysis treatment including reverse transcription on the treated cells, and respectively constructing a DNA library and an RNA library; and sequencing to obtain three-dimensional genomics information in a single cell. A high-throughput single-cell multi-omics three-dimensional genomics detection technology is designed, chromatin in cells is subjected to opening treatment through different methods, chromatin opening sites are enriched, the throughput of generated data is greatly improved, and the detection accuracy is greatly improved while the chromatin openness is detected in a single cell. The capability of detecting interaction between chromatin open sites and the gene expression capability are improved.

The invention provides a Chinese date modified acidic polysaccharide. Each 100 mg of the Chinese date modified acidic polysaccharide contains 90.26 mg + / -1.39 mg to 94.42 mg + / -1.83 mg of total polysaccharide, the total uronic acid content is 21.42 mg + / -1.01 mg to 50.72 mg + / -0.62 mg; total bound phenol: 0.65 mg of GAE + / -0.05 mg of GAE-1.21 mg of GAE + / -0.03 mg of GAE (equivalent weight of gallic acid). The invention also provides a preparation method and application of the jujube modified acidic polysaccharide. The jujube acidic polysaccharide is modified through a low-temperature alkaline deesterification technology, a directional enzyme digestion technology and a controllable partial acid hydrolysis technology, the in-vivo and in-vitro immunomodulatory activity of the jujube acidic polysaccharide before and after modification is compared, and the immunomodulatory activity of the jujube modified acidic polysaccharide obtained through modification is remarkably enhanced; especially, the jujube modified acidic polysaccharide with araban side chains removed and galactan side chains reduced has better immunocompetence.

The invention discloses application of a KASP molecular marker in identification of sweet melon peel color traits, and belongs to the technical field of molecular markers. The invention aims to promote gene localization and molecular-assisted breeding processes by utilizing molecular marker-assisted selective breeding on sweet melon pericarp color characters. The KASP molecular marker is a KASP marker RW2; the KASP marker RW2 is designed according to an SNP (Single Nucleotide Polymorphism) site at the position of Chr04: 526653. According to the application, the KASP molecular marker RW2 is used for performing genetic typing on the red-peel muskmelon and the white-peel muskmelon, the DNA of a target plant can be identified only through one-time PCR amplification, the whole detection process is simple and convenient, and complex steps such as enzyme digestion, electrophoresis and sequencing are not needed. Through the high-throughput genotyping system, a genotyping graph and a genotyping value can be rapidly obtained, so that rapid identification of the genotype of a target plant is realized. The KASP molecular marker RW2 provided by the invention can be more efficiently applied to the breeding work of sweet melon peel color germplasm.

A method for identifying an enzyme digestion site of a nucleic acid nickase is provided. The method uses mNGS sequencing to obtain nucleic acid sequence data before and after enzyme digestion, uses Bowite2 and samtools software to analyze second-generation sequencing data, and obtains the action site of the nickase according to the depth and analysis of single-base sequencing. The method does not need to clarify the sequence of the nucleic acid to be cut, takes a short time and low budget, and thus has a good application prospect.

The invention belongs to the technical field of proteomics, and particularly relates to a method for treating an extremely-micro tissue sample through a one-step method. The method comprises the following steps: 1, preparing a sample; 2, obtaining a peptide fragment: adding Mix Buffer into an ep tube, incubating after non-contact ultrasonic treatment, finally adding an acid solution to terminate enzyme digestion, and centrifuging to take a supernatant; the Mix Buffer is composed of rapid trypsin, CAA, TECP, DDM and a rapid trypsin buffer solution. According to the method, the Mix buffer is optimized, all steps of cracking, reduction, alkylation and enzyme digestion are combined into one step, the steps are simple, the cost is low, the sample preparation time is greatly saved, and a better proteomics test effect is achieved.

The invention relates to the field of analysis, in particular to an ovalbumin molecular weight analysis method and application thereof. The method for analyzing the molecular weight of the ovalbumin comprises the following steps: (a) performing denaturation treatment and cooling on an ovalbumin solution, and adding glycosidase for enzyme digestion to obtain an ovalbumin desugaring sample; (b) detecting and analyzing the ovalbumin desugaring sample by using liquid chromatography and mass spectrometry; and (c) carrying out molecular weight analysis according to detection data obtained by the mass spectrum. According to the method for analyzing the molecular weight of the ovalbumin, the desugaring step is simplified, and the desugaring effect is improved; the analysis time is short, the operation is simple, and only several hours are needed from sample pretreatment to result analysis completion.

The invention discloses a reconstructed respiratory syncytial virus F protein trimer, relates to the technical field of genetic engineering vaccines, and designs an RSV F protein trimer, a monomer of the trimer does not contain a p27 sequence, and a furin enzyme digestion peptide fragment aa110-aa136 is deleted; the F2 structural domain and the F1 structural domain are connected through a GS connexon; the invention also designs point mutation combinations, and the point mutation combinations can form an inter-chain disulfide bond, so that covalent connection of the trimer is realized, and the problem of depolymerization of the trimer is fundamentally solved; the trimer is stabilized in a pre-F state, a post-F sample structure does not exist, and an ELISA result shows that a neutralizing epitope is completely reserved; the trimer antigen structure is challenged by adopting a thermal acceleration experiment method, and a result shows that the purity of the trimer antigen reaches 95% or above after the trimer antigen is placed at 37 DEG C for 28 days, and the trimer antigen is very good in stability; when the trimer is used for preparing the vaccine, the standard required by laws and regulations can be met without freeze-drying.

The invention discloses an S-LAM-PCR library building method and application of the S-LAM-PCR library building method. According to the S-LAM-PCR library building method provided by the invention, the preference of enzyme digestion interruption is overcome by using ultrasonic interruption, the test period is shortened by using the PCR amplification kit and reducing the linear amplification cycle number, and the sensitivity of the method is improved by linear amplification and two rounds of nested PCR.

The invention discloses a circulating tumor DNA (deoxyribonucleic acid) standard substance as well as a preparation method and a valuing method thereof. The preparation method comprises the following steps: (1) culturing cells; (2) cell nucleus extraction; (3) carrying out enzyme digestion on cell nucleuses; (4) purifying the fragmented DNA; (5) sampling a sample, and quantifying the sample by nanodrop measurement; (6) sampling the length of a verification fragment; and (7) mixing different ctDNAs according to the finally obtained theoretical mutation abundance of 1%, 5% and 10% to prepare the standard substance. The valuing method comprises the following steps: carrying out digital PCR amplification on a sample through a double-labeled probe to obtain the copy number concentration of a mutant gene and a wild gene or the copy number concentration of a target gene and a reference gene, and calculating the gene mutation frequency and the amplification multiple. According to the method disclosed by the invention, the fragmented DNA with the length similar to that of the ctDNA is obtained; 330bp and large fragments can be removed through two rounds of purification of a paramagnetic particle method, and a single-fragment standard substance is obtained.

The invention discloses an anti-hepatoma drug screening cell model as well as a construction method and application thereof. The construction method of the model comprises the following steps: 1) fusing an EGFP (Enhanced Green Fluorescent Protein) gene to a C end of a Coilin gene, and constructing the fused gene to a CMV-MCS-EGFP-PGK-Puro lentiviral vector to obtain a virus vector for expressing Coilin-EGFP; 2) co-dyeing a virus vector and a virus packaging plasmid into a recipient cell for virus packaging, and centrifuging a collected cell culture solution containing the virus to obtain a virus supernatant for expressing the Coilinin-EGFP (Enhanced Green Fluorescent Protein); and (3) infecting the liver cancer cell line HCCLM3 by using the virus supernatant containing the expressed Coilinin-EGFP, and screening the cell strain capable of stably expressing the Coilinin-EGFP. According to the application, the obtained cell model is used for screening anti-hepatoma drugs. According to the method, the proper incision enzyme is selected, the enzyme digestion efficiency is high, the titer of the obtained virus vector expressing Coilinin-EGFP is high, the virus vector is applied to screening of anti-liver cancer drugs, positioning of Coilinin-EGFP agglomerates can be directly observed under a fluorescence microscope, the method is simple, visual and convenient to operate, and screening of compound small molecule drugs inhibiting cell nucleus positioning of Coilinin-EGFP is facilitated.

The invention discloses a construction method of a human epidermal organoid, and belongs to the technical field of organoid cell culture. The method is mainly used for optimizing a culture method of human epidermal organoid, and particularly relates to an efficient epidermal organoid culture technology based on a specific culture medium and an enzyme digestion system. According to the method, a # imgabs0 # human epidermal organ complete culture medium is adopted for preparation, and a basic culture medium and an additive are mixed to form the complete culture medium suitable for cell growth. The primary specimens are treated, and the steps of cleaning, digestion, cell collection and the like are included, so that the activity and functions of cells are ensured. Through accurate inoculation and culture conditions, the formation and growth of the organoid are promoted, and the culture in a proper environment is ensured, so that the formation efficiency of the organoid is improved. The organoid is subjected to passage and cryopreservation treatment, so that the activity and functional stability of the organoid in the long-term storage and use process are ensured.

The invention provides a method and system for discovering antioxidant peptides from food and agricultural by-products and outputting enzymolysis recommendation, and relates to the technical field of bioinformatics and artificial intelligence crossing. The method sequentially comprises the steps of protein sequence collection and standardization, candidate peptide fragment interception, prediction of antioxidant activity through fusion of sequence and structural characterization, generation of releasable peptide fragments through enzyme digestion simulation, and construction of comprehensive evaluation indexes based on activity probability and process parameters. And an optimal enzymolysis scheme is searched in an enzyme combination and reaction condition space through multi-objective optimization, so that the method is suitable for high-valued utilization of food industrial residues and agricultural wastes, and has the advantages of high throughput, high accuracy and high implementability.

The invention discloses a preparation method and application of siRNA (small interfering Ribonucleic Acid) for reducing silkworm moth wing area. The method comprises the following steps: extracting total RNA in a silkworm fat body, and carrying out reverse transcription to obtain first-chain cDNA; carrying out amplification by taking the silkworm Kurtz cDNA as a template to obtain silkworm Kurtz cDNA; carrying out PCR (Polymerase Chain Reaction) amplification by taking the pFLAG-BmKurtz as a template, and connecting a DNA (Deoxyribose Nucleic Acid) fragment with a vector after enzyme digestion to obtain Carrying out PCR (Polymerase Chain Reaction) amplification by taking the DNA as a template to obtain a reaction Mixing the reaction template with the mixed solution to react, and further treating to obtain BmKurtz siRNA; the BmKurtz siRNA is injected into the abdomen of pupated silkworm chrysalis, and the wing area of the pupated silkworm moth is reduced. The method is realized by regulating bombyx mori metabolism, genetic characteristics of bombyx mori are not changed, and research on substance and energy metabolism mechanisms of bombyx mori, regulation of bombyx mori physiological functions and the like is greatly promoted.

The invention discloses an EmAGO2 truncated recombinant protein, an echinococcosis multilocularis indirect ELISA (enzyme-linked immuno sorbent assay) diagnostic kit based on the truncated recombinant protein and a use method of the kit. The method comprises the following steps: designing a specific primer by selecting a sequence of 1-738bp at the N end of an Em-AGO2 gene, carrying out PCR (Polymerase Chain Reaction) amplification by taking reverse transcription echinococcosis multilocularis DNA (Deoxyribonucleic Acid) as a template, connecting an amplification product with a pCE2-TA / Blunt vector, and then transforming into a competent cell for culturing, so as to obtain a pCE2-T-EmAGO2 plasmid; respectively carrying out double enzyme digestion on the plasmid and an expression vector pET-28a (+), and connecting enzyme digestion fragments to obtain a recombinant protein expression plasmid pET-28a-EmAGO2; the preparation method comprises the following steps: taking EmAGO2 as a template, transforming the EmAGO2 into competent cells, selecting positive clones for induced expression, and carrying out ultrasonication treatment, separation and purification, concentration and desalination treatment on expressed thalli to obtain the EmAGO2 truncated recombinant protein. Furthermore, a mouse anti-EmAGO2 truncated recombinant protein and goat anti-rabbit IgG are respectively used as an antigen and an antibody to establish an indirect ELISA diagnostic kit for the echinococcosis multilocularis, and the kit has high sensitivity, specificity and good repeatability, and can be used for early diagnosis of the echinococcosis multilocularis.

The invention discloses a recombinant III-type human-like collagen as well as a coding gene and an expression method thereof, and the method comprises the following steps: S1, carrying out sequence optimization by taking a human III-type collagen sequence as a template to obtain a recombinant III-type human-like collagen gene sequence; designing PCR (Polymerase Chain Reaction) amplification upstream and downstream primers according to the gene sequence, and amplifying the gene sequence of the recombinant III type human-like collagen; s2, carrying out double enzyme digestion on the recombinant III type human-like collagen gene and a plasmid, and connecting to obtain a recombinant plasmid; s3, linearizing the recombinant plasmid, transforming the linearized recombinant plasmid into agrobacterium, and screening; and S4, infecting lettuce with agrobacterium, culturing to express protein, freezing the infected lettuce with liquid nitrogen, grinding, extracting total plant protein, and purifying to obtain the recombinant III-type human-like collagen. The recombinant III type human-like collagen is expressed through lettuce, so that the method is safe and efficient, and has sufficient market value and important practical significance.

The invention discloses a porcine circular RNA, a porcine circLDHC, an overexpression vector and a construction method and application thereof, and relates to the technical field of livestock molecular biology. The porcine circular RNA is formed by cyclizing 3rd to 6th exons of an LDHC gene, and the nucleotide sequence of the porcine circular RNA is as shown in SEQ ID NO: 1. The porcine circLDHC is obtained by carrying out reverse transcription on porcine circular RNA (Ribonucleic Acid). According to the invention, a specific amplification primer is designed for circLDHC, and the real existence of circLDHC is verified through PCR (Polymerase Chain Reaction) amplification, sequencing and RNase R enzyme digestion. Fluorescent quantitative PCR analysis finds that circLDHC is highly expressed in sexual mature testis tissues of pigs, the expression level of circLDHC is related to testis maturation, and meanwhile, the expression quantity of circLDHC in testis of 75-day-old Meishan boars is remarkably higher than that of circLDHC in testis of same-day-old Duroc boars (plt; 0.01) of the substrate. The invention further finds that the over-expressed circLDHC inhibits the proliferation of the porcine sertoli cells, promotes the maturation of the porcine sertoli cells, lays a foundation for further researching the start of the puberty period of the boars, and provides a new thought for analyzing the sexual precocious puberty mechanism of the boars and improving the reproductive performance.

The invention relates to polysaccharide-based composite hydrogel as well as a preparation method and application thereof. The composite hydrogel is prepared from the following raw materials: hydroformylated modified ortho-hydroxyl-containing polysaccharide, an extracellular matrix enzyme digestive juice, tannic acid, cannabidiol, neomycin sulfate and borneol, by compounding multiple components, the use performance of the hydrogel can be remarkably improved, and the hydrogel is particularly suitable for a harsh environment. The composite hydrogel disclosed by the invention has excellent mechanical properties, good biological safety, certain self-repairing performance and relatively high anti-inflammatory, antibacterial and antioxidant properties, and can be applied to skin or mucous membrane tissue wound repairing.

The invention discloses modified hydroxytyrosol, a cosmetic, a preparation method and application. The provided preparation method of the modified hydroxytyrosol comprises the following steps: in the presence of an acidic solvent, mixing hydroxytyrosol, lipase and fatty acid ester for reaction, centrifugally layering a reaction product, and collecting components of a middle layer to prepare the modified hydroxytyrosol. According to the provided preparation method of the modified hydroxytyrosol, lipase and fatty acid ester are subjected to an enzyme digestion reaction, an active hydroxyl group in a product obtained through the enzyme digestion reaction and hydroxytyrosol are subjected to a dehydration reaction, so that an ester group molecular chain is grafted into hydroxytyrosol, and a modified hydroxytyrosol product with an ester group is prepared. And the obtained modified hydroxytyrosol has relatively high fat solubility and oxidation resistance.

The invention relates to the technical field of gene engineering, in particular to a pineapple CRISPR / Cas9 gene editing vector as well as a construction method and application thereof. The invention relates to a pineapple CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / CRISPR associated protein 9) gene editing vector. Target sites Target1 and Target2 are designed according to a cDNA (complementary deoxyribonucleic acid) sequence of AcCWIN1; by taking a pMODB2312 carrier as a template, carrying out amplification by utilizing the primer for amplifying the target fragment to obtain the target fragment; a target fragment is inserted into a pMODB2312 carrier through double enzyme digestion, and a module B is obtained; the method comprises the following steps: cutting a pMODA1110 module, a pMODC0000 module, a pTRANS260d module and a B module by using an Aar I incision enzyme, and carrying out connection by using a T4 ligase, so as to obtain the CRISPR / Cas9 gene editing vector of the pineapple. The pineapple edited homozygous strain is successfully obtained according to the method, and a basis is provided for directional research of pineapples.