40 results about "Polyadenylic acid" patented technology

The invention provides a preparation method of a multivalent immunostimulation nanoformula. The preparation method comprises the steps of: (1) providing a nanogold aqueous solution; (2) modifying the 3' terminal of CpG ODN (oligodeoxynucleotides) by using polyadenylic acid to obtain CpG ODN-polyA; and (3) assembling the CpG ODN-polyA to nanogold to form the multivalent immunostimulation nanoformula. The invention further provides the multivalent immunostimulation nanoformula obtained by the preparation method and application of the multivalent immunostimulation nanoformula. Compared with the traditional CpG ODN, the multivalent immunostimulation nanoformula has very high stability, no influence on cell activity, high cellular uptake rate, low administration dosage, good immunostimulation activity and remarkable effect during administration of low dosage.

The invention relates to the technical field of molecular biology, in particular to eriocheir sinensis Crustin-1 gene clone and a recombination expression technology thereof. Crustin-1 gene of which the cDNA span is 821bp is cloned from eriocheir sinensis by using an expressed sequence tag (EST) technology and a 3' and 5' end rapid augmentation technology (RACE) and comprises a 315bp open reading frame, polyadenylic acid tailing signals and polyadenylic acid tails, 104 amino acids are coded, the length of a 5' non-coding region is 239bp, the length of a 3' non-coding region is 267bp, and the Crustin-1gene play an important role in the immune defense aspect of the eriocheir sinensis. The invention obtains eriocheir sinensis Crustin-1 protein by using the in-vitro recombination expression technology, the recombination protein has stronger bacteriostatic activity for gram-positive bacteria, the minimal inhibitory concentrations of the recombination protein for micrococcus luteus, bacillus subtilis, micrococcus tetragenus and bacillus thuringiensis are 0.12 mu m, 0.23 mu m, 0.46 mu m and 0.12 mu m respectively, and the recombination protein does not have obvious inhibiting function. The invention can lay the foundation for the disease control of the eriocheir sinensis, the gene assistant breeding and the development of feed additives.

The invention relates to the field of biotechnology, in particular to a mango ethylene receptor gene, which is named as MiETR1. Base sequences of the gene MiETR1 are as shown in SEQ ID NO:1 in a sequence table; the gene MiETR1 totally has 2570 base sequences including 113 3' untranslated regions, one polyadenylic acid tail and one opening decode regions with 2220 base sequences; an amino acid sequence of the gene MiETR1 is as shown in SEQ ID NO:2 in the sequence table. According to the mango ethylene receptor gene provided by the invention, the mango ethylene receptor gene MiETR1 is obtained through separating and cloning from mango, and an ethylene inhibitor is acted on the ethylene receptor gene MiETR1, so that an expression of the ethylene receptor gene MiETR1 can be remarkably inhibited, the ethylene release rate is reduced, the ethylene release peak is delayed, and the aim of delaying the ripening and senescence of the mango is achieved.

The invention discloses an expression vector expressing green fluorescent protein by fusion and a construction method and application thereof. The vector is in ring shape , comprising a procaryon replication origin, two eucaryon replication origins and selective marker genes, and two green fluorescent protein gene expression cassettes; the green fluorescent protein gene expression cassettes, from upstream to downstream, consist of two promoters with the same transcription direction, junction fragments, green fluorescent protein encoding genes and polyadenylic acid AATAAA; the green fluorescent protein encoding gene lacks an initiation codon. The junction fragment comprises an EcoRV recognition sequence; the first place and the second place of the green fluorescent protein encoding gene from the end of 5' overlaps the last two places from the end of 5' of the EcoRV recognition sequence; the junction fragment comprises or does not comprise the initiation codon; when the junction fragment comprises the initiation codon, the initiation codon has a space of 2+3n or 1+3n from the first place from the end of 5' of the EcoRV recognition sequence, wherein, n is a positive integer. The vector comprises a green fluorescent protein gene, and can only express the green fluorescent protein on the condition that the gene is correctly expressed, therefore, the interference due to the expression of fluorescence by an initial vector can be effectively excluded.

The invention discloses a method for preparing an mRNA and application of the mRNA in tumor treatment. The invention further discloses a nucleic acid molecule, which comprises a promoter, a transcribable segment A, a transcribable segment B, a transcribable segment C and a polyadenylic acid sequence in turn. Driven by the promoter, the transcribable segment A, the transcribable segment B, the transcribable segment C and the polyadenylic acid sequence can be co-transcribed to produce a common transcript; the segment A encodes a 5'-end untranslated region in the common transcript; the segment Cencodes a 3'-end untranslated region in the common transcript; the transcribable segment A is a DNA molecule shown as sequences 1-4; and the transcribable segment C is a DNA molecule shown as sequences 5-8. The mRNA molecule is obtained by in vitro transcription adopting the nucleic acid molecule as a template. The nucleic acid molecule or mRNA molecule provided can be used for preparing medicinesfor treating various tumors.

The present invention relates to an attenuated virus of a flavivirus and application of the attenuated virus. The attenuated virus comprises a polyadenylic acid (poly(A)) sequence, wherein the poly(A) is used for replacing a part of nucleotide sequence of a 3' untranslated region (3' UTR) of the yellow disease virus, so that the 3' untranslated region (3' UTR) of the attenuated yellow disease virus obtained after the part of nucleotide sequence of the yellow disease virus is replaced at least retains a 3'-end stem-loop region (3' SL). The attenuated virus has growth characteristics similar to those of a wild type virus, and is high in titer and good in hereditary stability. Moreover, the attenuation is fully attenuated, the safety is high, the generation of high-level neutralizing antibody titer can be induced by single immunization, the attack of high-dose wild-type viruses can be resisted, complete immune protection can be provided, and the attenuated vaccine strain can be used for preparing safe and effective attenuated vaccine strains.

The invention discloses an rAAV vector for expressing an antibody IgG1 and application of the rAAV vector. The ITR core expression element of the rAAV vector is composed of a broad-spectrum strong promoter CMV, an IgG1 heavy chain secretion signal peptide sequence, an IgG1 heavy chain coding region, a connecting polypeptide coding sequence, an IgG1 light chain secretion signal peptide sequence, an IgG1 light chain coding region and a human serum albumin polyadenylic acid sequence. The heavy chain variable region (VH) and the light chain variable region (VL) can be conveniently and quickly operated through the restriction enzyme cutting sites on the rAAV vector. The antibody IgG1-rAAV obtained by packaging is used for infecting HEK293T cells or C57 mice and the like cultured in vitro, and various antibodies IgG1 with different antigen recognition activities can be expressed.

The invention discloses a swine promoter protein expression vector and a construction method and application thereof. The expression vector is annular and comprises a pronucleus replication origin, an eukaryon replication origin, a selection marker gene and an exogenous gene expression cassette, wherein the exogenous gene expression cassette consists of a transcription promoter, a junction fragment and a polyadenylic acid tailing signal sequentially from upstream to downstream; and the transcription promoter is an MLP6 promoter. The expression vector can efficiently express exogenous proteinsin a swine cell, contains the eukaryon replication origin and can be proliferated greatly in a cell of an expression T antigen, so that protein expression level is raised. A recombinant vector of an expression exogenous gene is constructed by homologous recombination, so that the conventional steps such as enzyme cutting, linking and the like are avoided and 2 to 3 steps are saved compared with the conventional linking process. A vector recombination process for constructing the expression exogenous gene by using the vector of the invention is simple, convenient, rapid, economical and efficient and the construction of the expression vectors of a large number of exogenous genes can be finished rapidly.

The invention provides a prawn virus expression system for delivering and expressing an exogenous gene in a prawn cell, which comprises a recombinant plasmid for expressing the exogenous gene in the prawn cell, the recombinant plasmid comprises genome DNA (deoxyribonucleic acid) of prawn IHHNV virus, a promoter PH (potential of hydrogen) of insect polyhedrosis protein gene, a polyclone restriction enzyme cutting site MCS (multi-cloning restriction enzyme cutting site) for inserting an exogenous gene and an SV40 polyadenylic acid signal pA. The expression system further comprises an insect baculovirus recombinant expression plasmid and an insect Sf9 packaging cell, wherein the gene of the WSSV envelope protein VP28 is inserted into the insect baculovirus recombinant expression plasmid. The prawn virus expression system provided by the invention can efficiently express exogenous genes in prawn cells, and all plasmid DNA genetic information in the expression system is free out of chromosomes of the prawn cells and cannot be integrated into genomes of the prawn cells, so that the biological safety of the system is improved.

The invention relates to the field of molecular biology, in particular to a cell-free translation system, method and product. The translation system comprises mRNA of 5'cap and 3 'polyadenylic acid, an amino acid mixture, an energy molecule and a mixture of mRNA translation enzyme sources; the energy molecule comprises phosphocreatine, ATP (adenosine triphosphate) and guanosine triphosphate (GTP). Through mRNA transcription, in-vitro transcription and gel electrophoresis analysis, the translation system can rapidly obtain a large amount of target proteins and has the following characteristics: an extracellular translation technology is adopted to overcome the problem that a prokaryotic cell expression part gene cannot be expressed; the problems of long expression cycle and low efficiency of eukaryotic cells are solved by adopting an extracellular translation technology; the problems of low activity and poor efficiency of the original extracellular translation technology are solved by adopting a strategy of adding energy micromolecules such as phosphocreatine; and the stability, the receptor binding speed and the overall translation speed are enhanced by adopting mRNA of 5 ' cap and 3' polyadenylic acid.

The present invention relates to an oncolytic virus and use thereof, the oncolytic virus comprising an attenuated flavivirus, the attenuated flavivirus comprising a polyadenylic acid (poly (A)) sequence, wherein the poly (A) sequence is used for replacing a part of nucleotide sequence of a 3' untranslated region (3' UTR) of the yellow disease virus, so that the 3' untranslated region (3' UTR) of the attenuated yellow disease virus obtained after the part of nucleotide sequence of the yellow disease virus is replaced at least retains a 3'-end stem-loop region (3' SL). In-vitro cell experiments prove that the oncolytic virus disclosed by the invention can infect and kill various tumor cells, but has small influence on normal cells; and a xenotransplantation tumor model proves that the oncolytic virus disclosed by the invention can inhibit the growth of various tumor cells in vivo, and a safe and efficient anti-cancer therapeutic agent with small toxic and side effects can be provided for clinical tumor treatment.