41 results about "Full length genome" patented technology

The present invention provides a method for replicating efficiently an RNA containing fulllength HCV genomic sequence and a method for producing HCV virus particles containing fulllength HCV replicon RNA or fulllength HCV genomic RNA by using a cell culture system. Further, the present invention relates to a method for producing hepatitis C virus particles which comprises culturing a cell, into which a replicon RNA comprising a nucleotide sequence comprising a fulllength genomic RNA sequence of hepatitis C virus of the genotype 2a, at least one selectable marker gene and / or at least one reporter gene and at least one IRES sequence or the fulllength genomic RNA of hepatitis C virus of the genotype 2a is introduced, and generating virus particles in the culture medium. Still further the present invention relates also to a hepatitis C vaccine and an antibody against hepatitis C virus particles.

An allergen-free transgenic peanut seed is produced by recombinant methods. Peanut plants are transformed with multiple copies of each of the allergen genes, or fragments thereof, to suppress gene expression and allergen protein production. Alternatively, peanut plants are transformed with peanut allergen antisense genes introduced into the peanut genome as antisense fragments, sense fragments, or combinations of both antisense and sense fragments. Peanut transgenes are under the control of the 35S promoter, or the promoter of the Ara h2 gene to produce antisense RNAs, sense RNAs, and double-stranded RNAs for suppressing allergen protein production in peanut plants. A full length genomic clone for allergen Ara h2 is isolated and sequenced. The ORF is 622 nucleotides long. The predicted encoded protein is 207 amino acids long and includes a putative transit peptide of 21 residues. One polyadenilation signal is identified at position 951. Six additional stop codons are observed. A promoter region was revealed containing a putative TATA box located at position −72. Homologous regions were identified between Ara h2, h6, and h7, and between Ara h3 and h4, and between Ara h1P41B and Ara h1P17. The homologous regions will be used for the screening of peanut genomic library to isolate all peanut allergen genes and for down-regulation and silencing of multiple peanut allergen genes.

The invention relates to construction and application of SHIV (simian / human immunodeficiency virus) infectious clone. Firstly, the overall length genome of a virulent strain SHIV-KB9 embedded with a B subtype env gene is cloned to a low copy number carrier pBR322, the obtained plasmid SHIV-KB9 / pBR322 is taken as a genome skeleton, the env gene of Chinese main prevalent strain CRF08_BC of HIV-1 is used for substituting for corresponding gene segments so as to obtain the infectious clone SHIV-KBQJ-12, the replacing env gene is from the infectious clone strain QJ001 of HIV-1CRF08_BC and comprises the whole intracellular region, a transmembrane region and part of an extracellular region of gp120 and gp41, and the position of the replacing env gene, corresponding to the position of the nucleotide of a standard reference strain HXB2, is 6103-8249. The construction is simple and convenient to operate, has high success rate, can obtain the infectious clone in a short time, and can be used for establishing an SHIV / Chinese rhesus infection model.

The present invention provides a rabies virus CTN strain whole genome cDNA infectious clone, as well as a method of using reverse genetics technology to save the active rabies virus. The present inventive method includes: using the single restriction enzyme sites in the CTN strain of rabies virus full-length genome to divide the full-length virus into four segments to amplify, simultaneously using the green fluorescent protein gene to substitute 423bp base at the pseudo-gene point, cloning in the pVAX-R expression vector, to construct a recombinant full-length plasmid of the virus genome; performing PCR amplification of nucleoprotein, phosphoprotein, glycoprotein and transcription large protein gene sequence of the CTN strain of the rabies virus, and cloning the amplified fragment into a vector pVAX1, to constitute four auxiliary plasmids in the reverse heredity system; and then using the five plasmids to jointly transfect cells, to rescue the rabies virus. The present invention successfully saves the rabies virus, and has a great significance in the study of the pathogenic mechanism of rabies virus, development of new vaccines, viral vectors and the like areas.

The invention relates to a preparation method of a HPV full-length genome quality control product, an amplification primer and a detection reagent. Wherein, the preparation method of the HPV full-length genome quality control product comprises the following steps: (1) obtaining a DNA template; (2) using amplification primers to perform PCR amplification; (3) using HD Cloning connection technology to connect the target DNA fragment to the carrier ; (4) introducing the plasmid into competent cells, and then extracting the plasmid; (5) diluting the plasmid with human genome. The preparation method of the human papillomavirus full-length genome quality control product of the present invention can successfully obtain the HPV gene detection quality control product that has no biological infection risk type, can simulate clinical samples, can be produced in large quantities, has a controllable concentration, and is stably stored. Has good applicability.

The invention discloses a restructured infectious cloning plasmid of a low virulent strain of a Japanese encephalitis virus, which contains a full-length genome cDNA sequence of the low virulent strain 10S3 of the Japanese encephalitis virus, or a full-length cDNA sequence of a marked cleavage site introduced in the full-length genome cDNA sequence of the low virulent strain 10S3 of the Japanese encephalitis virus. The invention also discloses an infectious cloning low virulent strain of the Japanese encephalitis virus. The low virulent strain is a cloning strain of the low virulent strain 10S3 of the Japanese encephalitis virus, and the full-length genome cDNA sequence of the low virulent strain 10S3 is shown in SEQ ID NO.1. The restructured infectious cloning plasmid of the low virulent strain of the Japanese encephalitis virus and the infectious cloning low virulent strain of the Japanese encephalitis virus rescued from cells can provide complete immunoprotection for immunized animals resisting Japanese encephalitis viruses, thereby extremely facilitating the prevention and control of Japanese encephalitis viruses.

The invention discloses an adenovirus vector and its application in preparing HCV (hepatitis C virus) cells and mouse models. The invention provides a recombinant adenovirus vector which is obtained by inserting nucleotides shown by the sequence 1 in the sequence table in the adenovirus vector HDAd (helper-dependent adenovirus). The experiments prove that the invention provides a third generation adenovirus vector of a recombinant HCV 2a type full-length genome, the prepared adenovirus can be used for preparing HCV whole-genome cell models or mouse models, and the obtained models can be used for drug screening of anti-HCV drugs, so as to be used for HCV vaccine evaluation and researches of the pathogenic mechanism.

Belonging to the technical field of biological genetic engineering, the invention in particular relates to a hordeum vulgare calmodulin gene HvCAM1 and salt tolerant application thereof. The hordeum vulgare calmodulin gene HvCAM1 has a nucleotide sequence shown as SEQ ID No:1 and a full-length genome sequence 2124 bp. According to the invention, firstly a hordeum vulgare calmodulin coded gene is obtained by cloning and is named as HvCaM1, and the transcript cDNA full-length nucleotide sequence is shown as SEQ ID No:2, and the full-length cDNA sequence is 728 bp. The invention also utilizes RNAi interference method to silence the HvCAM1 gene successfully in hordeum vulgare Golden Promise, also utilizes the material to verify the negative regulation effect of HvCAM1 gene under salt stress can effectively enhance the salt tolerance of hordeum vulgare. At the same time, overexpression of the HvCAM1 gene in hordeum vulgare Golden Promise cannot enhance the salt tolerance of hordeum vulgare.

The invention discloses a GII.4 type norovirus genome amplification primer and an amplification method. In the present invention, six pairs of primers are used as the upstream and downstream primers of the amplification primers, and the RNA of GII.4 type norovirus is used as the template to carry out RT-PCR amplification to obtain amplification products respectively, and then perform nucleic acid sequence sequencing on the amplification products , and then spliced ​​and compared to obtain the full-length sequence of the GII.4 norovirus genome. According to the main popular genotypes of GII.4 Norovirus, the present invention designs a "4+1+1" segmentation amplification strategy according to the three open reading frames contained in the genome, and designs corresponding amplification according to the conserved region Primer; it can effectively amplify the whole genome sequence of GII.4 type norovirus. The invention can be widely used in medical and health care, inspection and quarantine, etc., which have norovirus detection requirements and corresponding scientific research fields.