152 results about "Protein translation" patented technology

The invention provides a preparation method of a yeast cell extracting solution, the yeast cell extracting solution and a method for synthesizing protein in an in vitro system by virtue of the yeast cell extracting solution, as well as a kit containing the yeast cell extracting solution. The kit, which is more convenient than a conventional method, is applicable to protein synthesis in the in vitro system; transformation, culture and crushing can be prevented; a great amount of using time and cost can be saved; various types of proteins can be expressed, and influence of protein toxicity can be prevented; and a plurality of protein complexes can be expressed, without the use of a high temperature at 37 DEG C. In addition, as a raw material adopted by the invention, yeast cell is simple to cultivate, convenient to operate, rapid to propagate and relatively low in cost; a prepared yeast extract has a capacity of modifying translated protein, the yeast extract is suitable for large-scale preparation and has an advantage of industrial production; and the yeast cell extract, which is prepared into freeze-dried powder by virtue of a vacuum freeze-drying method and is preserved at low temperature or high temperature, is convenient to transport.

The invention provides a nucleic acid construct for endogenously expressing RNA polymerase in cells, and particularly discovers a novel nucleic acid construct capable of greatly enhancing protein translation efficiency. The nucleic acid construct consists of a promoter with specific promotion intensity (such as RNR2, ADH1, GAPDH, TEF1, PGK1 and SED1) and coding sequences of proteins of RNAP (suchas T7RNAP, T3RNAP, T4RNAP and T5RNAP), if the nucleic acid construct of the invention is applied into a yeast extracorporeal protein synthesis system, the activity of synthesized luciferase is quite high relative to light unit value (RLU), and the effect can be the same as the effect of T7RNAP added exogenously. The effect can reach up to 6.6* 107.

The invention provides a DNA element for high-throughput in vitro protein synthesis, and specifically discloses a novel DNA element capable of greatly enhancing protein translation efficiency. According to the present invention, with the application of the DNA element in a yeast in-vitro protein synthesis system, the relative light unit value of the activity of the synthesized luciferase is enhanced by about 22.3 times compared to the DNA element containing only the omega sequence.

The invention provides preparation of an eIF4G and Pab1 fusion protein in deficiency of different structural domains and application of the fusion protein to improvement of protein synthesis. Particularly, the fusion protein in deficiency of different structural domains is capable of changing in-vitro protein translation efficiency, wherein RNA1 and/or PABP structural domain deficiency of an eIF4Gelement can remarkably improve protein expression. In addition, the invention provides a preparation method of the infusion protein and a corresponding in-vitro protein synthesis system and method.

The invention provides a protein synthesis system for increasing the expression amount of a foreign protein and an application method of the protein synthesis system. Specifically, a poly-nucleotide sequence or vector constructed by a strong promoter sequence is inserted in front of a nucleotide sequence for encoding an eIF4E binding protein, over-expression of eIF4E binding proteins such as Eap1and p20 can be achieved, and foreign protein synthesis is implemented by using the improved cell lysis buffer. The invention provides the protein synthesis system for increasing the expression amountof the foreign protein, and by adopting the synthesis system provided by the invention, the efficiency of protein translation can be remarkably improved.

The invention provides a detecting method of post-translational protein modification. The method comprises the following steps: calculating spectrogram difference vectors among all spectrograms by means of peptide chromatogram retaining time and peptide quality in experimental tandem mass spectrum data of protein samples; establishing candidate modification mass intervals probably containing modification mass; estimating mixed distribution of the spectrogram difference vectors on each candidate modification mass interval, and calculating the standard deviation of each distribution inside the mixed distribution so as to determine distributions inside the candidate modification mass intervals caused by post-translational protein modification according to the standard deviation; calculating the average value of the distributions caused by post-translational protein modification; obtaining an accurate mass experimental value of the post-translational protein modification according to the mass component of the average value; and obtaining the influence of post-translational protein modification on peptide chromatogram retaining time according to the retaining time component of the average value. The detecting method has the advantages of high efficiency, accuracy and robustness.

The present invention provides a cyclic RNA preferable for carrying out rotary protein translation in which translation domains other than that of the target protein are sufficiently short and translation efficiency is high, and a method for producing protein that uses this cyclic RNA as template. More specifically, the present invention provides a cyclic RNA that encodes a protein, has a full-length number of bases that is equal to or greater than 102 and is a multiple of 3, has at least one start codon, does not have a stop codon in the same reading frame as the start codon, and does not contain an internal ribosome entry site (IRES). In addition, the present invention provides a method for producing protein in a eukaryotic cell expression system that consists of using the aforementioned cyclic RNA as template and expressing a protein encoded by that cyclic RNA.

Provided are methods and devices for identifying pathogens based on protein translation from ribosomes isolated from the pathogens.

Disclosed is a method for monitoring early treatment response of a cancer treatment comprising measuring by magnetic resonance spectroscopy (MRS), for example, proton MRS, the amount of Choline present in the tissue adjoining or surrounding the cancerous tissue before and after treatment; the treatment comprises administration of an angiogenesis inhibitor, for example, a VEGF inhibitor, whereby a decrease in the amount of Choline after treatment is indicative of a positive response. The decrease in the amount of Choline represents the decrease in the internal cell membrane as a result of down regulation of the organelles and their secretory granules and their transport vesicles. Disclosed also is a method for determining effectiveness of an angiogenesis inhibitor in the treatment of cancer. Also disclosed are methods of monitoring early treatment response in diseases where an angiogenesis effector, i.e., an inhibitor or promoter of angiogenesis, is employed. Also disclosed is a method for monitoring protein translation related to angiogenesis.

The invention provides a method for analyzing data of prokaryotic proteogenomics rapidly and automatically. Users only need to provide mass spectral data and a corresponding database file and set a simple retrieval parameter; retrieval of data of proteogenomics can be completed; simultaneously, a user defined data retrieval result can also be compatible; therefore, the identification coverage rate of the data of proteogenomics is increased; in the method disclosed by the invention, library search engines of different algorithms are integrated in advance; disadvantages of a single retrieval method are made up; a user defined library search result can also be compatible; the method has good compatibility; the peptide fragment identification coverage rate is increased to the most extent; in the method disclosed by the invention, functional annotation of new genes is automatically completed; large-scale identification of protein post-translational modification and analysis of non-labelled quantitative proteomics are realized for the first time; and automatic and rapid deep analysis of the data of proteogenomics is really realized.

The invention relates to the enrichment and purification of phosphopeptide in phosphoproteome, in particular to the application of ZrO2 in the enrichment and purification of phosphopeptide. ZrO2 nano material shows high specificity, selectivity, binding capacity and sensitivity at a phosphopeptide section; meanwhile, the affinity of the ZrO2 nano material to the specificity of phosphopeptide ensures that the material can be used in phosphoproteome during protein post-translational modification. Due to the specificity combination of ZrO2 with the phosphopeptide section in complex proteolysis sample, large-scale identification of phosphorylated protein and determination of phosphorylation sites can be realized through combining with MALDI-TOF MS and nano-LC MS/MS.

The invention discloses a replicative HBV (Hepatitis B Virus) vector carrying a foreign gene and a recombinant HBV generated after transfection and a corresponding preparation method and an application. The vector separates overlaying genes C and P on an HBV genome by a molecular cloning technique based on originally expressed HBV plasmid to respectively form an integral opened reading frame where a protein translation starting sequence or a protease enzyme cutting site is inserted to respectively guide expression of foreign gene and gene P. The replicative HBV vector carrying the foreign gene transiently transfecting hepatoma carcinoma cell secretes the recombinant HBV. The recombinant HBV prepared from the HBV vector transfection cells provided by the invention can express the foreign gene and maintain the replicative and infecting capacity. The invention is suitable for constructing an HBV chronic infection animal model, an HBV cell model with cccDNA stably and automatically replicated and a traceable HBV strain, researching a molecular mechanism of HBV infection, replication, packaging and the like, and screening anti-HBV novel medicines.

The current invention provides a method for detecting the presence of a genomic or proteomic precursor(s) within a sample, wherein the precursor(s) provide an indication of the presence or the capability of expression modulation of various other proteins which may, either directly or indirectly, provide an indication, promote, and/or be responsible for the post translational modification of the PCNA.

The invention discloses a brown planthopper protein translation elongation factor NlEF1gamma, an encoded protein, synthesized dsRNA, and an application thereof in controlling brown planthopper pest diseases. The gene performs an important function in maintaining the normal growth and development processes of brown planthoppers. When the function is inhibited, brown planthopper nymph growth is slower, brown planthopper nymph size is smaller, and brown planthopper nymph dies because it couldn't develop into adult. When the function of the gene in a brown planthopper female adult is inhibited, its ovaries stop developing and it couldn't spawn normally. According to the invention, brown planthoppers are injected and fed with the dsRNA of the gene fragment. With both means the brown planthopper death rates reach 70% and brown planthopper spawning amounts are 0. Sequence and data bases are provided for establishing novel strategies for controlling pests with an RNA interference technology. The application has significant lethal and ovarian development inhibition effects against brown planthoppers. Therefore, the gene can be sued as an effective target for controlling the pest with an RNAi technology.