16results about How to "Genetic stability" patented technology

The invention provides a construction method of a non-small cell lung cancer histone H1.3 arginine methylation point mutation cell model. The construction method comprises the following steps: designing mutation primers H1.3 R80A-F and H1.3 R80A-R; the method comprises the following steps: by taking a pCDH-HA-H1.3-Flag plasmid as a template, carrying out PCR (Polymerase Chain Reaction) amplification by adopting a mutation primer, digesting a product by DMT enzyme, and converting a competent cell to obtain a mutant plasmid; and co-transfecting the mutant plasmid and a helper plasmid to a packaging cell, collecting a virus solution, filtering, infecting an A549 cell, adding Polybrene to assist infection, culturing, and screening a stably transfected cell strain by using puromycin to obtain the recombinant plasmid. The invention also provides application of the mutant cell model obtained by the construction method. According to the invention, the blank of histone H1.3 methylation research is filled, the 80th arginine is clear as a core modification site, and the established model provides a new tool for lung cancer mechanism research and drug research and development.

This application relates to a species of Pseudomonas aeruginosa. bpsR Genes, BpsR proteins and their applications, engineered bacteria, and drugs, among which bpsR The nucleotide sequence of the gene is shown in SEQ ID No. 1. This invention... bpsR Genes, through regulation in Pseudomonas aeruginosa bpsR Gene expression can regulate the virulence factors of Pseudomonas aeruginosa, such as biofilm production, pyocyanin production, motility, and elastase secretion levels. Furthermore, BpsR protein can be used as a new target for the development of drugs to prevent and treat Pseudomonas aeruginosa, which has important clinical significance for the prevention and control of Pseudomonas aeruginosa infection.

The invention provides a sheep embryo fibroblast line and application thereof, and belongs to the technical field of cell biology. The name of the sheep embryo fibroblast line is sheep embryo fibroblast OEF, the sheep embryo fibroblast line is preserved in China Center for Type Culture Collection, the address is Wuhan University, Wuhan, China, the preservation date is January 26, 2022, and the preservation number is CCTCC NO: C202212. The sheep embryo fibroblast line can be used for separation and culture of bovine and sheep pox viridae viruses such as LSDV, GTPV, SPPV and ORFV, has the characteristics of short proliferation time and high virus titer, and can avoid the risk that other viruses are polluted by viruses generated by primary cell separation.

The invention provides a skin organoid HPV infection model based on a human embryonic stem cell source and a construction method and application thereof, and relates to the technical field of molecular biology, the construction method comprises the following steps: integrating an HPV genome to an AAVS1 site of genome DNA of a human embryonic stem cell to obtain the human embryonic stem cell carrying the HPV genome, and performing induced differentiation and activation of the HPV genome to obtain the human embryonic stem cell carrying the HPV genome. A skin-like organ HPV infection model is formed, stable heredity and controllable expression of an HPV genome are achieved, virus particle assembly is achieved through induced differentiation, establishment of a virus replication environment, activation of HPV early gene expression, driving of HPV genome replication and induction of late gene expression, and the natural infection state of HPV is simulated. The technical problem that in the prior art, an infection model cannot simulate the life cycle of HPV dependent terminal differentiation is solved, and an irreplaceable technical platform is provided for research of an HPV pathogenic mechanism and research and development of antiviral drugs and vaccines.

The invention discloses a DNA structure for saccharomyces cerevisiae single-cell barcode labeling and chromatin state conjoint analysis and application thereof, and belongs to the technical field of molecular biology and single-cell multi-omics analysis. From the 5'end to the 3 'end, a fluorescent protein gene, a Tn5ME structure, a reverse transcription primer binding region, a CYC1 terminator fragment and a linearized shuttle plasmid skeleton are subjected to homologous recombination, and a pCM carrier is obtained; after NdeI / BamHI enzyme digestion, oligonucleotides with random sequences, which are subjected to annealing, amplification, Klenow filling-in and enzyme digestion, are connected, and transformation extraction is carried out, so that the DNA structure for saccharomyces cerevisiae single-cell barcode labeling and chromatin state combined analysis is obtained. On the basis of the constructed DNA structure, ATAC-seq and a bar code reverse transcription method are combined, important data related to the transcription state and chromatin state of a single cell can be obtained at the same time through bar code information, single cell pedigree tracking of the saccharomyces cerevisiae is achieved, and research on single cell epigenomics of the saccharomyces cerevisiae and other microorganisms is promoted.

The application discloses a deletion mutant of a rice LHCB5 gene and application thereof, and belongs to the field of gene editing. Through gene editing system, the 3'UTR region of the rice LHCB5 gene is precisely edited, and a 40bp homozygous deletion mutant of +16 to +55bp downstream of the termination codon TAA is obtained. The mutant can be stably inherited, and the growth phenotype is not significantly different from that of the wild type. Meanwhile, the resistance to rice blast and bacterial leaf blight is significantly improved, and the length of the disease spot is significantly reduced. The application first discloses the disease resistance regulation function of the 3'UTR of the LHCB5 gene, realizes the improvement of the disease resistance through the editing of the non-coding region, avoids the safety and agronomic trait risks brought by the transgene and the coding region editing, and the obtained mutant can be directly used for the breeding of the rice broad-spectrum disease resistance, and has important application value.

The present application relates to a high-yield Morchella MZ-12 and its cultivation method, belonging to the technical field of edible mushroom cultivation. The present application uses chemical reagent N-methyl-N'-nitro-N-nitrosoguanidine (NTG) mutagenesis method to first obtain a Morchella MZ-12, which is preserved in the China General Microbiological Culture Collection Center on March 23, 2023, with the preservation number of CGMCC No.40394; the strain has a shortest mushroom growth time shortened by 10 days compared with the original Morchella strain, has a higher yield, has a strong resistance to Streptomyces, has genetic stability, has a more thick and elastic flesh compared with the known Morchella varieties, has a more rich aroma, and has a better commodity nature.

This invention discloses a mutant attenuated strain of avian reovirus genotype II, wherein the σC protein amino acid sequence of the mutant strain includes mutations at positions 40, 45, 111, 180, 244, 273, and 300. Due to this mutation, compared to the parent wild-type strain, the pathogenicity to SPF chickens is significantly reduced, it does not cause persistent footpad swelling, and the viral load and shedding levels in tissues are extremely low, without affecting weight gain. Immunopotency evaluation shows that this mutant attenuated strain can induce high levels of specific antibodies, effectively controlling clinical symptoms, viral load in tissues, and shedding. This invention provides a safe and highly effective attenuated vaccine candidate strain for the prevention and control of avian reovirus genotype II, filling a corresponding technological gap.

The invention belongs to the technical field of plant genetic engineering, and particularly relates to application of a PcWVD2 gene in enhancing the dwarfing ability of pear plants. The CDS (Coding Sequence) of the PcWVD2 gene is as shown in SEQ ID NO. 1. The amino acid sequence coded by the PcWVD2 gene is as shown in SEQ ID NO. 2. The invention reveals that the PcWVD2 gene has the function of regulating and controlling the dwarf character of the pear plant, the dwarf ability of the pear can be obviously enhanced by over-expressing the gene, the dwarf ability shows that the plant height is reduced, the internode is shortened and the root length is shortened, and an important candidate gene is provided for dwarf close planting cultivation of the pear tree. According to the method for enhancing the dwarfing ability of the pear plant, the endogenous dwarfing gene PcWVD2 is directly overexpressed by the pear plant through genetic transformation, the stably inherited dwarfing character can be realized, the dependence on dwarfing rootstock can be gotten rid of, and the cultivation management cost can be reduced.

This invention belongs to the field of trastuzumab technology, specifically relating to a Nod2 gene knockout CHO cell line, its preparation method, and its applications. The Nod2 gene in the knockout CHO cells is lost. The CHO cells stably express trastuzumab before Nod2 gene knockout. At the same cell density, the CHO cell line shows a 1.22-1.55-fold increase in trastuzumab expression. This invention knocks out the Nod2 gene in CHO cells using a CRISPR / Cas9 system, and also provides a method for constructing the cell line, a method for producing trastuzumab using the cell line, and applications of the cell line. The Nod2 gene knockout cell line significantly increases trastuzumab expression in suspension culture while effectively reducing the aggregation level of the target protein. Furthermore, the high-yield trastuzumab trait is stably inherited and does not significantly inhibit normal cell growth, providing a new stable platform for efficient trastuzumab production and possessing significant industrial application value.

The invention relates to a yellow flammulina velutipes C53 with a preservation number of GDMCC No.67334. The yellow flammulina velutipes C53 has the advantages of high yield, short growth cycle, more edible parts of stipes, difficulty in browning of stipe bases, long preservation time and stable inheritance, and is used for cultivation and breeding of flammulina velutipes. The invention also relates to an identification method of the flammulina velutipes C53, which comprises the following steps: by taking the DNA of the flammulina velutipes mycelium to be detected as a template, carrying out PCR (Polymerase Chain Reaction) amplification by adopting the primer combination 1 and / or the primer combination 2, and detecting whether an amplification product has a band with a specific size so as to identify the C53. The screening method is simple to operate, and detection can be performed only by using mycelia; the detection time is only 2-3 hours, so that the breeding efficiency is improved, and the cost is greatly saved.

This invention discloses a high-glutathione-producing engineered Escherichia coli strain and its applications, belonging to the field of biotechnology; the engineered Escherichia coli strain is named ZMGSH01F, with accession number CCTCC M 20252236; the engineered Escherichia coli strain is based on Escherichia coli BL21(DE3) as the starting strain, with gene clusters knocked out. ptsH‐ ptsI‐crr And integrate at this site glf‐glk Fusion genes, knockout gene clusters gsiA‐gsiB‐gsiC Integrate the endogenous glutathione transporter gene at this site. cycDC Knockout of γ-glutamyltransferase gene ggt Simultaneously expressing the recombinant plasmid pRSFDuet- gshAB K177E‑E275K‑V513A - ppk2 The engineered Escherichia coli strain can efficiently produce glutathione through fermentation culture using glucose, monosodium glutamate, cysteine, and glycine as substrates.

This invention provides an L-homoserine-producing strain, its construction method, and its applications. The strain does not contain plasmids. E.coli W3110, as a chassis strain, had the relA and thrB genes knocked out, and heterologously expressed genes derived from... Clostridium acetobutylicum The gapC gene and the aspB gene derived from Bacillus subtilis were overexpressed, along with ppc, asd, and thrA. fbr ,rhtA,pntAB,spoT fbr Genes; the strain described above can effectively improve the L-homoserine synthesis efficiency by knocking out genes of the L-homoserine degradation pathway, enhancing the expression of key enzymes in the pathway, optimizing the L-homoserine transport system, increasing the reducing power NADPH content, and enhancing the ability to withstand amino acid starvation. It is genetically stable, requires no addition of resistance substances, has high acid production efficiency, and has good L-homoserine synthesis capacity.

The application discloses application of a CsKIC gene and a protein coded by the same in regulation of resistance to cucumber powdery mildew, wherein a nucleotide sequence of the CsKIC gene is shown as SEQ ID NO:1, and an amino acid sequence of the protein is shown as SEQ ID NO:2. By using a VIGS gene silencing technology, it is found that silencing of the CsKIC gene increases the disease resistance of cucumber to powdery mildew, thereby verifying that the CsKIC gene is a recessive disease resistance gene related to cucumber powdery mildew. The application discovers a new function of the CsKIC gene in relation to the resistance of cucumber to powdery mildew, provides a new selection and target for breeding of cucumber powdery mildew resistance, and lays a foundation for development of new cucumber germplasm resources.

This invention discloses the construction and application of a xanthan gum cell factory with short fermentation cycle, high potency, and high resistance to oxidative stress. Using *Xanthomonas aeruginosa* XC1 as the starting strain, this invention employs a multi-step chromosome integration strategy to achieve the synergistic effect of three major modules: "enhancing quorum sensing signals, strengthening the rate-limiting synthesis step, and improving antioxidant performance." The resulting engineered strain EG-Y6 showed a 66-fold increase in intracellular DSF levels compared to the original strain XC1, significantly advancing the efficient xanthan gum synthesis period and shortening the fermentation cycle from 72 hours to 48 hours. The xanthan gum fermentation potency reached 26.9 g / L, a significant increase of 33.2% compared to the basic strain XC1. In the later stages of fermentation, the intracellular glutathione (GSH) content was 2.22 times higher than that of the basic strain XC1, while the reactive oxygen species (ROS) level decreased by 57%.

The application discloses a genetic hyperparathyroidism animal model and a construction method and application thereof, and relates to the technical field of animal models.The point mutation animal model constructed by the application not only provides a reliable basis for verifying pathogenicity of GCM2 mutation, but also lays a repeatable and generalizable technical foundation for clarifying a molecular mechanism of genetic hyperparathyroidism, screening potential treatment targets, and developing related pharmaceutical research.