82 results about "Antigen Gene" patented technology

The invention relates to the technical field of cell engineering, and particularly discloses a construction method and application of a sika deer immortalized renal epithelial cell line, and the construction method comprises the following steps: taking renal epithelial primary cells from healthy sika deer renal cortex tissues to obtain primary renal epithelial cells; treating the primary renal epithelial cells with trypsin to obtain a cell suspension; inoculating the cell suspension into a complete culture medium, and transfecting with lentivirus loaded with SV40 large T antigen genes to obtain transfected cells; 3 [mu] g / mL puromycin is applied to the transfected cells for selective culture, so that non-transfected cells are eliminated, and drug-resistant cells are obtained; carrying out continuous passage on the drug-resistant cells for at least 30 generations to obtain the immortalized renal epithelial cell line of the sika deer; according to the invention, through a specific action mechanism (inhibiting a p53 / pRb pathway and blocking cell cycle exit) of the SV40 large T antigen, a proliferation limit caused by inhibition of telomerase activity of primary cells of the cervidae animals is overcome, and a cell resource library capable of realizing continuous passage is established.

The invention provides human trichomonad antigen gene protein glyceraldehyde-3-phosphate dehydrogenase, and particularly relates to application of the protein in immunological detection of human trichomonad infection, the protein can be used for immunological detection of human trichomonad infection, and a human trichomonad detection kit is prepared. The GAPDH protein is used as a detection antigen for indirect ELISA detection, and the method has the characteristics of high sensitivity and specificity.

The invention relates to the technical field of biological genetic engineering, in particular to construction of recombinant lactobacillus plantarum of an African swine fever virus antigen, and the method comprises the following steps: constructing a lactobacillus plantarum shuttle vector containing different ASFV antigen genes; and referring to a genome sequence of a Chinese isolate CADCHN09 of ASFV disclosed in GenBank. The method has the advantages that an inducer is not needed, antigen protein is efficiently expressed, the dendritic cell (DC) binding peptide for enhancing immune cell uptake and processing is subjected to fusion expression, the oral immunization mode is simple, mucosal immunity is stimulated, and ASFV infection is effectively prevented and controlled; probiotics, namely lactobacillus plantarum, which naturally exist in animal intestinal tracts and can be colonized and copied in digestive tracts for a long time are used as a carrier; key epitopes of proteins such as nucleocapsid protein p72, outer envelope protein CD2v, inner envelope protein p54, p22, pE199L and pE248R, which play an important role in adsorption, internalization, endocytosis and membrane fusion of ASFV, are preferably fused with dendritic cell binding peptide and then are displayed on the surface of lactobacillus plantarum.

The invention relates to the field of intelligent medical treatment, in particular to a new antigen screening method, equipment and a program product. Comprising the following steps: S1, acquiring tumor slice data and serum data of a patient; s2, performing data processing on the tumor slice data and the serum data to obtain an HLA / MHC pseudo sequence, antigen peptide data and an antigen gene expression level; the data processing comprises data sequencing, HLA / MHC typing, somatic mutation calculation, fusion gene calculation and gene expression profile calculation; s3, screening new antigens based on the HLA / MHC pseudo sequence, the antigen peptide data and the antigen gene expression level data to obtain the new antigens, the application can perform new antigen screening, and has good clinical value.

The invention discloses a surface antigen gene StSAG1 of Sarcocystis tenella, the nucleotide sequence of the surface antigen gene StSAG1 is as shown in SEQ ID NO: 1, and according to the analysis result of protein antigenicity and antigen epitope, the 686 bp-1175 bp segment of the StSAG1 gene is selected to construct a recombinant protein expression vector and engineering bacteria; the antigen disclosed by the invention has very strong antigenicity and specificity, and can be subjected to effective antigen-antibody reaction with an antibody generated in serum after sheep (Ovis aries) is infected with sarcosporidium tenella, so that diagnosis is realized; low-titer antibodies can be detected two weeks after infection, and the detection window period is obviously advanced; by adopting the ELISA detection method, a visual result can be directly obtained. The method breaks through the limitation of an infection window period, has the advantages of low cost, high sensitivity, simplicity and convenience in operation, no need of expensive instruments and analysis and the like, is suitable for field detection in primary laboratories and pastures, and is beneficial to industrial production and market popularization and application.

The invention relates to the technical field of immunodetection, and discloses a bovine nodular skin disease virus chimeric antigen and application thereof in an ELISA (enzyme-linked immuno sorbent assay) detection kit. According to the chimeric antigen, a gene of a dominant region of a bovine nodular skin disease virus antigen is selected to design a chimeric antigen gene, meanwhile, sequence optimization is performed, a chemical synthesis method is used for synthesizing a coding gene of the chimeric antigen of the bovine nodular skin disease virus, and then the chimeric antigen is obtained in a prokaryotic expression mode. An ELISA antibody detection kit developed on the basis of the chimeric antigen has extremely high sensitivity and specificity, can rapidly and specifically detect bovine nodular skin disease virus antibodies in serum, is suitable for monitoring and checking the nodular skin diseases of cattle in large-scale breeding, and has relatively high practical value.

The present disclosure relates to methods for characterizing cancer testis antigen burden (“CTAB”), for predicting cancer survival outcomes using CTAB analysis, and for recommending and / or treating cancer using CTAB analysis. Particularly, aspects are directed to measuring expression of a panel of cancer testis antigen (CTA) gene markers in a tumor sample from a subject, determining a CTAB based on the measured expression of the CTA gene markers, classifying, by comparing to a reference CTAB score from the same cancer or tumor type, the CTAB score as a high-CTAB score or a low-CTAB score, and identifying the subject as responsive to a treatment wherein (i) the high-CTAB score is indicative of responsiveness to the treatment, or (ii) the low-CTAB score is indicative of responsiveness to the treatment.

This invention discloses a recombinant adenovirus vaccine targeting the Tp0326 antigen and its preparation method, comprising a recombinant adenovirus vector and the Tp0326 antigen expressed therein. The recombinant adenovirus vector uses a replication-defective human adenovirus type 5 (Ad5) as a backbone, inserting the full-length Tp0326 antigen gene of *Treponema pallidum* into the adenovirus shuttle vector pshuttle-IRES-rGFP-1, and obtaining it through homologous recombination with the adenovirus backbone plasmid pAdEasy-1. This invention, using a replication-defective Ad5 as a vector, provides a recombinant adenovirus vaccine with high safety and efficient expression of the full-length Tp0326 antigen. The expressed antigen has a natural conformation, retains the ECL4 immunodominant epitope, and is more likely to induce a specific immune response, resulting in stronger immunogenicity. The preparation method involves constructing a recombinant adenovirus vector through homologous recombination, packaging it in HEK293T cells, and purifying it by CsCl density gradient centrifugation to obtain a high-purity, high-titer recombinant adenovirus vaccine suitable for large-scale production. The recombinant adenovirus vaccine can be administered via intramuscular or intranasal injection.

The invention relates to a full-length sequence amplification primer, an amplification method and a three-generation sequencing method of an HLA-I antigen gene, and belongs to the field of gene detection. The HLA-I antigen gene is subjected to PCR (Polymerase Chain Reaction) amplification by using 8 pairs of characteristic amplification primers. And each gene can be subjected to high-yield specific amplification on a full-length sequence containing complete intron and exon regions only through one round of amplification and enrichment, and 7.8 Kb long-fragment genes can be captured once at most. The experimental deviation caused by simultaneous amplification of a single gene by using a plurality of pairs of segmented primers and the indirect error caused by gene splicing are effectively avoided. The method comprises the following steps: carrying out quality inspection and purification on amplicon PCR (Polymerase Chain Reaction) products, mixing samples, constructing an HLA-I type antigen gene library, carrying out accurate and complete sequencing on the full-length sequence of the HLA-I type antigen gene by adopting a three-generation PacBio Sequel II platform, and ensuring that the Hifi reads base accuracy can reach 99% or above. The reading of HLA-I type antigen gene full-length sequence variation information and the haplotype analysis at a high resolution level can be realized.

The present invention relates to the field of medicine, and specifically discloses an influenza virus vaccine fusion protein, which is a fusion protein formed by connecting an HA antigen fragment encoded by an influenza HA antigen gene and an M2 extracellular region fragment (M2e; Membrane Protein (M2) Extracellular Region (M2e) in series with an Fc fragment encoded by a human immunoglobulin gene in series. The fusion protein of the present invention has higher antigenicity than a single HA antigen or a single M2 extracellular region fragment antigen, and the Fc fragment can improve protein purification efficiency and prolong the half-life of the antigen in vivo. Therefore, the influenza vaccine provided by the present invention has great advantages and great application value.

The invention discloses a sgRNA sequence for targeted knockout of an O antigen gene cluster OL101 and application of the sgRNA sequence. The invention relates to an sgRNA sequence for targeted knockout of an O antigen gene cluster OL101. The sequence comprises oagen-sgRNA-F as shown in SEQ ID NO. 1 and oagen-sgRNA-R as shown in SEQ ID NO. 2. The invention relates to a CRISPR / Cas9 and lambda Red plasmid vector system for targeted knockout of an O antigen gene cluster OL101, the plasmid vector system is a sgRNA sequence for targeted knockout of the O antigen gene cluster in claim 1 and a recombinant expression plasmid vector of lambda Red and CRISPR / cas9 vectors, and the sequence of the plasmid vector system is as shown in SEQ ID NO.3. According to the invention, the Klebsiella pneumoniae OL101 can be specifically knocked out in a targeted manner, so that an OL101 knocked-out strain is obtained, and the specific functions of the Klebsiella pneumoniae O antigen OL101 can be clarified. According to the system for targeted knockout of the Klebsiella pneumoniae O antigen OL101, O antigen gene clusters of strains can be effectively knocked out, the gene knockout period is short, and the method is simple.

The new alpharetrovirus-based particles are suitable for high efficiency of transiently transducing animal cells. e.g. human or murine cells, and which efficiently introduce coding and non-coding RNA contained in the alpharetrovirus-based particles into target cells The alpharetrovirus-based particles also provide for high efficiency of the activity and / or integrity of the RNA that is introduced into the animal cells, as the particles protect the incoming RNA from degradation during entry. The transferred RNA can be of non-coding nature (e.g. single guide (sg) RNA. short-hairpin (sh) RNA. micro RNA. or long non-coding (Inc) RNA. or the RNA may encode proteins or peptides. e.g. receptors. transcription factors. cellular enzymes, antigens for use in vaccination, gene / protein therapy and / or gene editing nucleases, recombinases and transposases.

The invention relates to a recombinant rabies virus. The recombinant rabies virus is obtained by inserting an antigen gene of feline calicivirus and an antigen gene of feline herpes virus into a genome of a rabies virus attenuated strain. The triple inactivated vaccine for preventing and treating the feline calicivirus disease, the feline herpes virus disease and / or the rabies is prepared based on the recombinant rabies virus, and after the triple inactivated vaccine is used for immunizing cats, neutralizing antibodies aiming at the feline calicivirus disease, the feline herpes virus disease and the rabies virus can be effectively induced and generated; the simultaneous immunization on the three viruses is realized.

The invention provides two specific diagnostic antigens for chicken infected with dog head roundworm and an ELISA diagnostic kit thereof, and belongs to the technical field of biological immunodetection, and the two specific diagnostic antigens are a recombinant protein GAPDH3 and a recombinant protein dim-1; the nucleotide sequence of the specific diagnosis antigen gene GAPDH3 is as shown in SEQ ID NO.1, and the amino acid sequence of the specific diagnosis antigen gene GAPDH3 is as shown in SEQ ID NO.3; the nucleotide sequence of the specific diagnosis antigen gene dim-1 is as shown in SEQ ID NO.2, and the amino acid sequence of the specific diagnosis antigen gene dim-1 is as shown in SEQ ID NO.4. The kit is used for detecting the specific antibody of the dog head roundworm in chicken serum, detection signals can be displayed more clearly in the detection process, misjudgment caused by non-specific reaction is reduced, and the detection accuracy and reliability are improved. The condition of the specific antibody of the dog head roundworm in chicken serum can be more accurately judged, and a more reliable basis is provided for diagnosis of chicken infection of the dog head roundworm.

The application belongs to the technical field of biology, and discloses a preparation method of a Salmonella typhimurium flagellin and Senecavirus antigen fusion protein. The application selects Escherichia coli Rosetta (DE3) as a host, fuses a Salmonella typhimurium flagellin gene and a Senecavirus antigen gene, optimizes codons, and then clones the fusion protein into a pET-28a (+) vector, so that the obtained fusion protein can keep the adjuvant activity of the flagellin, improve the solubility and expression level of VP2, and the yield can reach 40 mg / L, thereby greatly reducing the expression and purification process cost. The fusion protein provided by the application can activate the TLR5 path and activate mouse macrophages, and the production process is simple and low in cost, so that the application has important significance for the research and production of Senecavirus subunit vaccines.

The invention discloses a human immortalized fallopian tube fibroblast strain as well as a construction method and application thereof, and belongs to the technical field of cell immortalization construction. The preparation method comprises the following steps: firstly, separating, culturing and purifying fibroblasts from human fallopian tube tissues by virtue of an enzyme digestion method, identifying markers of the fibroblasts, then introducing a simian virus 40 large T antigen gene into primary fibroblasts by virtue of a lentiviral vector, and screening by virtue of puromycin to obtain a cell strain capable of stably expressing SV40T. Finally, the human immortalized fallopian tube fibroblast which can be stably passaged is constructed. The cell strain has the characteristics of strong multiplication capacity and high hereditary stability, and the cells are not obviously aged along with the increase of the number of generations of the cells, which indicates that the human immortalized fallopian tube fibroblast line is successfully established. According to the invention, the problem that the in-vitro culture life of primary fibroblasts is limited is solved, and a reliable in-vitro cell model is provided for research in the fields of fallopian tube related diseases, ovarian cancer origin, tumor microenvironment and the like.

The invention relates to the technical field of mucosal immunity and immunogenicity collaborative optimization, in particular to a preparation method of a mucosal immune type mycoplasma bovis live vector vaccine, attenuated mycoplasma bovis is used as a vector, an inducer X containing an FRET modified nucleic acid aptamer is combined with CRISPR-Cas12a for modification, a virulence gene is knocked out, and a mucosal targeting gene fused with CTB is inserted; carrying out deep codon optimization on the antigen gene, fusing the antigen gene with the glycosylation immunopotentiation peptide, and carrying out novel enzyme digestion connection to construct a recombinant vector; and intelligently fermenting in an improved culture medium, centrifuging, resuspending by using a special protective agent, and freeze-drying to obtain the vaccine. According to the preparation method, the mucous membrane targeting and immunogenicity of the vaccine are improved, and the production process is optimized; mucosal immunity can be quickly stimulated, various immune reactions are activated, the vaccine activity and quality are guaranteed, and the mycoplasmosis bovis is efficiently prevented.

The present application relates to the technical field of novel coronavirus, and particularly relates to a novel coronavirus trivalent DNA vaccine and a preparation method. The trivalent DNA vaccine takes a recombinant plasmid carrying a nucleotide sequence as shown in SEQ ID NO: 1 or 2 as an active ingredient. The trivalent DNA vaccine provided in the present application can express a fusion antigen protein of three kinds of antigen genes of the novel coronavirus in eukaryotic cells, and produce serum IgG antibodies against the three kinds of antigens of the novel coronavirus. In addition, the novel coronavirus trivalent DNA vaccine provided in the present application can immunize mice to produce T cells secreting various cytokines, and has a stronger T cell immune protection effect.

The invention discloses an HMGCR antigen gene, an HMGCR recombinant antigen gene, a recombinant antigen, a reagent for detecting an anti-HMGCR antibody as well as a preparation method, a detection method and application of the reagent, the HMGCR antigen gene is obtained by modifying a gene for coding HMGCR protein by adopting a codon optimization technology, and the HMGCR recombinant antigen gene is obtained by modifying the optimized HMGCR antigen gene. The nucleotide sequence of HMGCR is optimized by using a codon optimization tool, and the sequence after ATG is subjected to recombination transformation, so that the expression quantity of HMGCR protein is improved; through further recombination transformation, the expression quantity of the HMGCR antigen is further improved, and the expression level of protein can be greatly improved by applying the sequence which is easier to express. The kit can be used for detecting the anti-HMGCR antibody in human body fluid, the detection specificity and sensitivity are improved, and compared with WB, the kit has higher sensitivity and specificity, and the specificity and sensitivity reach 100%.

The application discloses a recombinant bacillus Calmette-Guerin (BCG) strain and application of the recombinant BCG strain in preparation of a tumor vaccine, wherein the recombinant BCG strain comprises an HPV16 E7 antigen gene with a nucleotide sequence as shown in SEQ ID NO:1; a cytolysin ClyA gene is further fused with the HPV16 E7 to be expressed, and a rBCG-ClyA+E7 recombinant strain is constructed; the recombinant BCG strain rBCG-E7 or rBCG-ClyA+E7 is applied in preparation of a vaccine for treating cervical cancer solid tumors; the anti-tumor effect is verified in a solid tumor model through a subcutaneous injection mode; the ClyA significantly improves the expression efficiency of an exogenous antigen in the BCG, and simultaneously enhances the presentation efficiency of an antigen presenting cell; the recombinant vaccine effectively activates an anti-tumor immune response, and does not cause obvious toxic side effects, thereby providing a safe and effective candidate strategy for immunotherapy of cervical cancer and other HPV related solid tumors.

The application provides application of rotavirus non-structural protein 4 in improving immune function of recombinant rotavirus subunit vaccine, wherein the application is to express the rotavirus non-structural protein 4 gene and rotavirus antigen gene in different surface rings on a norovirus P particle to obtain a recombinant protein, and the recombinant protein is the recombinant rotavirus subunit vaccine. The application improves the immune function of the recombinant rotavirus subunit vaccine by using the NSP4 protein, and parallel expression of the NSP4 gene and the VP8 antigen gene or the NSP4 antigen gene can improve the NSP4 antigen and the VP8 antigen specific IgG antibody titer, improve the neutralization ability of the VP8 antigen to the rotavirus, and improve the protection of the NSP4 antigen and the VP8 antigen to the diarrhea mice induced by the rotavirus SA11 strain.

The invention relates to a mycobacterium abscessus and marine mycobacterium vaccine and a preparation method thereof, the mycobacterium abscessus and marine mycobacterium vaccine comprises a recombinant plasmid based on an antigen protein gene, the recombinant plasmid is used for stably expressing cell wall antigen protein, and the recombinant plasmid is used for stably expressing cell wall antigen protein. The recombinant plasmid sequentially contains a resistance screening gene Kan, an escherichia coli replicon oriE, a promoter hsp60, an antigen protein gene MMAR1296 and a gene tag Flag-tag in the clockwise direction, wherein the DNA (deoxyribonucleic acid) sequence of the promoter hsp60 is as shown in SEQ ID No.1; the recombinant plasmid sequentially contains a resistance screening gene Kan, an escherichia coli replicon oriE, a promoter hsp60, an antigen protein gene MMAR1296 and a gene tag Flag-tag in the clockwise direction; the sequence of the antigen protein gene MMAR1296 is as shown in SEQ ID No. 2; and the DNA (Deoxyribose Nucleic Acid) sequence of the gene tag Flag-tag is as shown in SEQ ID No. 3. According to the invention, cell wall antigen protein MMAR1296 from marine mycobacterium is displayed on the surface of a mycobacterium smegmatis Msmeg vaccine carrier without any toxicity, a recombinant strain and a vaccine thereof are constructed, and subcellular component separation and western-blot experiments prove that the antigen protein MMAR1296 is successfully displayed on the surface of mycobacterium smegmatis MC2155; balb / c mouse in-vivo experiments prove that the immune effect of the recombinant mycobacterium smegmatis vaccine with the MMAR1296 antigen protein displayed on the surface is far superior to that of a bacillus calmette guerin vaccine BCG.

The present invention discloses a recombinant Bacillus subtilis and a construction method and application thereof, belonging to the field of biological gene engineering technology. The recombinant Bacillus subtilis is constructed by introducing an expression vector containing a Bacillus subtilis spore capsid protein gene and a porcine reproductive and respiratory syndrome virus antigen gene; the capsid protein gene is CotB or CotC; the antigen gene is a fusion gene ORF56 of ORF5 and ORF6. The present invention also discloses the application of the recombinant Bacillus subtilis in the preparation of a vaccine for the prevention and treatment of porcine reproductive and respiratory syndrome. The recombinant Bacillus subtilis prepared by the present invention has anti-PRRSV virus activity, can produce humoral immunity and cellular immune response in mice, not only provides a new idea for the research and development of vaccines, but also provides a strong technical support for the healthy breeding of pigs.

According to one embodiment, a lipid particle for producing a CAR antigen-expressing cell derived from a CD209 positive cell is provided. The lipid particle includes a lipid membrane having a lumen, and a nucleic acid containing a CAR antigen gene being contained in the lumen.

The invention provides a pertussis mRNA vaccine, and belongs to the technical field of biology. Antigens of the pertussis mRNA vaccine comprise pertussis toxin and filamentous hemagglutinin, a gene sequence of the pertussis toxin comprises gS1 and gC180, ninth arginine of amino acid sequences corresponding to the gS1 and the gC180 mutates into lysine, and 129th glutamic acid of the amino acid sequences corresponding to the gS1 and the gC180 mutates into glycine. A signal peptide of a human Igk light chain is added to the upstream of the amino acid sequence to replace a natural bacterial signal peptide; the gene sequence of the filamentous hemagglutinin comprises an FHA456 gene, an FHA373 gene and an FHA233 gene. According to the invention, the sequences for coding the antigen genes gS1, gC180, FHA456, FHA373 and FHA233 are optimized, so that an organism can be efficiently induced to generate immune response.

Vibrio parahaemolyticus serotype O10 variants are important food-borne pathogenic bacteria with extremely high separation rate in recent years in China. The invention provides a specific oligonucleotide sequence for detecting vibrio parahaemolyticus serotype O10 variants and application of the specific oligonucleotide sequence. The oligonucleotide is DNA fragments screened from IS1 genes and orf9 genes in vibrio parahaemolyticus serotype O10 variant O antigen gene clusters respectively. The invention also establishes a set of PCR detection method for detecting vibrio parahaemolyticus serotype O10 variants by using the oligonucleotide, establishes a technology for detecting the vibrio parahaemolyticus serotype O10 variants by using a molecular biology means for the first time, and has important significance on identification of the important food-borne pathogenic bacteria and epidemiological monitoring.

The invention discloses a listeria monocytogenes multi-epitope fusion antigen, a listeria monocytogenes multi-epitope fusion antigen-based nano-particle and application of the listeria monocytogenes multi-epitope fusion antigen-based nano-particle. Amino acid sequences of important antigen protein gene encoding proteins of LM antigens are analyzed, dominant antigen epitopes are screened out and connected in series according to a certain permutation and combination, a multi-epitope fusion antigen gene LM-MeAg is constructed and synthesized, and the listeria monocytogenes multi-epitope fusion antigen-based nano-particle is obtained. And further expressing the LM multi-epitope fusion antigen LM-MeAg gene in escherichia coli, and constructing a pT-LM-MeAg5 recombinant plasmid through a molecular cloning technology. According to the invention, dominant antigen epitopes of important antigens LLO, ActA, InlB and MPL of LM are assembled together in series according to a certain permutation and combination through nucleotides for coding flexible peptide amino acids, an LM multi-epitope fusion antigen gene is constructed, and an LM-MeAg5-PLGA nano-particle molecule with strong immunogenicity of the LM multi-epitope fusion antigen protein is further prepared. The method can be used for researching, developing and preparing efficient and safe anti-LM vaccines or drugs.

The application belongs to the technical field of biotechnology, and provides a porcine deltacoronavirus recombinant DNA vaccine, a construction method and application thereof, the porcine deltacoronavirus recombinant DNA vaccine comprising a porcine deltacoronavirus NTD antigen gene or a porcine deltacoronavirus RBD antigen gene; wherein the nucleotide sequence of the porcine deltacoronavirus NTD antigen gene is shown as SEQ ID NO. 1, and the nucleotide sequence of the porcine deltacoronavirus RBD antigen gene is shown as SEQ ID NO. 2. The application firstly fuses the immunogenic gene fragments NTD and RBD of PDCoV with IgG1 Fc fragments respectively, constructs a recombinant DNA vaccine, and evaluates the safety, immunogenicity and protective efficacy of the recombinant DNA vaccine through experiments, and verifies that the PDCoV recombinant DNA vaccine has good effects in preventing porcine deltacoronavirus infection.

The invention discloses a method for recovering adipogenic gene expression and adipogenic capacity of bovine immortalized adipocytes based on siRNA (small interfering Ribonucleic Acid). In order to solve the problems that bovine immortalized adipocytes established by overexpression of SV40 T-antigen genes are inhibited in adipogenesis related gene expression and abnormal in adipogenesis ability, the invention inhibits the expression of SV40 virus T-antigen in bovine adipocytes by designing, screening and identifying specific efficient interfering siRNA (small interfering ribonucleic acid) aiming at the SV40 virus T-antigen. Therefore, the inhibition effect of SV40 on adipogenic gene expression is relieved, the expression level and the adipogenic capacity of the adipogenic gene in the bovine immortalized adipocytes are recovered, and the method plays a key role in breaking through the bottleneck problem of developing species-specific adipodifferentiation mechanism research by utilizing the bovine immortalized adipocytes and getting rid of the current situation of developing research by depending on a mode animal cell line for a long time.

A method of early detection of Type 1 diabetes uses a quantitative assay to measure biomarkers of autoimmune diseases at specific short regions of a gene sequence. The assay uses amplicon deep sequencing to quantify individual variants of the ERV Group Antigen (Gag) gene associated with an overactive immune response.