11 results about "Chaperone Gene" patented technology

The invention relates to the technical field of biology, in particular to a recombinant pichia pastoris engineering bacterium, a construction method of the recombinant pichia pastoris engineering bacterium and application of the recombinant pichia pastoris engineering bacterium in secretory expression of ovotransferrin. The heterologous expression of ovotransferrin is effectively realized by optimizing ovotransferrin genes and utilizing molecular chaperone genes to strengthen the regulation and control capability of hosts, and meanwhile, by optimizing signal peptides and constructing high-copy recombinant pichia pastoris expression strains, the low-copy expression condition is verified and compared by utilizing fermentation expression, so that the heterologous expression of ovotransferrin is realized. According to the high-copy recombinant pichia pastoris engineering bacteria constructed by the invention, the ovotransferrin expression quantity is obviously improved and reaches 140 mg / L, the secretory expression efficiency of the ovotransferrin is effectively improved, and theoretical guidance is provided for heterologous expression and industrial production of the ovotransferrin.

The invention discloses a method for synthesizing 11 alpha hydroxyprogesterone by using cytochrome P450 enzyme recombinant bacteria, and belongs to the technical field of biology. The method comprises the following steps: firstly, constructing CYP219A1 gene and reduction chaperone Pdx / Pdr gene expression vectors from neosphingosinearomaphilus, and transferring the CYP219A1 gene and reduction chaperone Pdx / Pdr gene expression vectors into escherichia coli to obtain whole-cell transformed recombinant bacteria; and after culture, feeding a substrate progesterone by using a resting cell method for biotransformation to obtain a 11 alpha-hydroxyprogesterone product. According to the CYP219A1 mutant strain provided by the invention, the conversion rate of progesterone is as high as 99%, and the 11 alpha site selectivity is 74%.

The application belongs to the technical field of biology, and discloses a ferritin expression and purification method in bacillus subtilis, wherein the food safety level strain bacillus subtilis WB600 is used as a host, and the synergistic regulation of expression element (promoter, signal peptide) screening, bottom strain (dltD gene knockout, csaA molecular chaperone gene overexpression) modification and induction condition (induction time, temperature, IPTG concentration) optimization is used to successfully realize the high-efficiency secretory expression of Japanese capsule prawn ferritin (MjFer) and H-2 type soybean ferritin (H-2). The transmission electron microscope and dynamic light scattering analysis prove that the two proteins both retain the typical cage structure of ferritin, and the iron oxidation precipitation and reduction release experiment proves that the two proteins retain the basic iron oxidation precipitation and reduction release characteristics of ferritin, which is helpful for the large-scale production of ferritin in the food industry.

The present invention relates to a method for detecting a partner gene, which comprises extracting total RNA of a sample, reverse transcribing to obtain cDNA, then fragmenting into 150-250 pb fragments, repairing and adding A at the ends, connecting sequencing adapters, and obtaining a prelibrary; blocking the prelibrary with a blocking blocker, then hybridizing a specific probe set with the prelibrary to obtain captured fragments, enriching and purifying the fragments by PCR, and then performing quality inspection to obtain a sequencing library; sequencing the sequencing library using second-generation sequencing to obtain gene fusion information. The method of the present invention can still maintain a high capture efficiency while using fewer probes and a lower amount of sequencing data, obtain more positive reads than traditional capture, reduce probe synthesis and sequencing costs, and improve detection sensitivity.

The application discloses a recombinant Pichia pastoris genetic engineering bacterium with high yield of lactoferrin, and simultaneously satisfies the following three modifications: overexpression of a bovine lactoferrin expression gene, mutation of an alpha mating factor signal peptide at position E86 to F, and overexpression of a PDI molecular chaperone gene. The application also discloses a construction method of the recombinant Pichia pastoris genetic engineering bacterium, and application of the recombinant Pichia pastoris genetic engineering bacterium in production of bovine lactoferrin. The 5L fermenter test result of the bacterium strain shows that the yield of exogenous protein is significantly improved, and is increased from 10.04 mg / L in a flask fermentation to 95.78 mg / L, nearly 10 times. The application is based on the Pichia pastoris system and is not related to complex culture condition change, and is low in cost and easy to enlarge. The method is not only suitable for production of bovine lactoferrin, but also provides a new modification idea for high-efficiency expression of other heterologous proteins in the yeast system.

This invention relates to a nanopore sequencing detection method for sarcoma EWS-ETS fusion genes based on single-end primers. It designs an amplification strategy combining a 3' gene-specific single primer with a universal primer, leveraging the long read advantage of nanopore sequencing. A complete detection workflow is constructed, encompassing sample pretreatment, nucleic acid extraction, single-end primer amplification, nanopore library construction and sequencing, and fusion gene identification. The clinical application pathway is also clearly defined. This invention eliminates the need for prior knowledge of the fusion partner gene, significantly improving the detection sensitivity of rare EWS-ETS fusion genes. It offers high breakpoint localization accuracy and standardized procedures, enabling efficient screening and confirmation of sarcoma fusion genes, providing reliable technical support for accurate diagnosis, prognostic assessment, and targeted therapy of sarcoma.

The present invention relates to the compound of formula (1a) wherein p is 1 or 2, R1-R4 are hydrogen atom or the like, Ring A is cycloalkylene or the like, L is single bond or the like, and R is methyl or the like, or a pharmaceutically acceptable salt thereof, which has an anticancer effect by inhibiting the binding between a MLL fusion protein that is fused with AF4, AF9, or the like, which is a representative fusion partner gene causing MLL leukemia, and menin.

The invention discloses a probe set, a kit and a method for detecting blood tumor genes, the probe set comprises a plurality of probes, 1 / 3-2 / 3 of the two adjacent probes are overlapped during targeted detection, and each basic group in a targeted detection area is covered by the two probes. The invention provides a 2 * Tiling designed probe group, and each basic group in a detection area is covered by two primers, so that the capture efficiency is improved by about 20% compared with a conventional imbricated designed probe. For fusion detection, not only common driver genes but also partner genes are contained. Probe design is carried out in a mode of laying probes at a breakpoint and an exon boundary region, and the detection sensitivity of fusion variation is remarkably improved. In addition, the probe density is adjusted according to the GC content, and the high-GC region adopts a'high-density coverage 'strategy, so that the coverage degree is remarkably improved. Aiming at the characteristic that the blood tumor fusion partner gene is highly homologous, the optimized sequence is used as a reference sequence to detect the IGH / IGJ / IGK region, so that the problem of multiple comparison is avoided.

A method for screening αβ-TCR mutants, the method comprising the construction of a phagemid. In the construction, a gene encoding an A1 polypeptide and a gene encoding B1 polypeptide are inserted into the 3′ ends of genes encoding the constant regions of the α chain and the β chain of TCR respectively, and a ribosome binding site and a molecular chaperone gene are integrated into a phage vector, wherein the amino acid sequence of the A1 polypeptide is as shown in SEQ ID NO: 1 or is represented by the variant of the A1 polypeptide; the amino acid sequence of the B1 polypeptide is as shown in SEQ ID NO: 2 or is represented by the variant of the B1 polypeptide; and the variants at least retain the function of the polypeptides before mutation. The A1 peptide and the B1 peptide are used for promoting the correct pairing and folding of the αβ heterodimer TCR, so that the efficiency of displaying the correctly folded TCR on the surface of a phage is improved. The method can efficiently complete the screening of αβ heterodimer TCR mutants.

The present invention relates to the compound of formula (1a) wherein a-d and p are 1 or 2, R1-R4 are hydrogen atom or the like, and R18 is —CF3 or the like, or a pharmaceutically acceptable salt thereof, which has an anticancer effect by inhibiting the binding between a MLL fusion protein that is fused with AF4, AF9, or the like, which is a representative fusion partner gene causing MLL leukemia, and menin.

The invention discloses a prediction model construction method for RUNX1 rearrangement leukemia prognosis and sensitive drugs, and belongs to the technical field of biomedicine. The invention discloses an objective rule that the RUNX1 fusion gene causes abnormally high expression of the partner gene through a'promoter exchange 'mechanism for the first time, establishes a prognosis layering prediction model based on the partner gene pan-cancer high expression risk ratio (HR), divides the RUNX1 fusion gene into a high risk group and a low risk group, and establishes a prognosis layering prediction model in TARGET-AML (n = 3, 4, 5, 6, 7, 8, 8, 9, 10, 11, 12, 13, 13, 13, 13, 13, 13, 13, 13, 13, 13, 13, 13, 13, 13, 13, 157) and CNpAML (n = 149) queues verify their predicted value for patient lifetime and minimal residual focus level. Meanwhile, a prediction model of RUNX1 fusion gene sensitive drugs based on partner gene drug sensitivity characteristics is developed. The clinical problems that RUNX1 rearrangement leukemia risk stratification is incomplete, evaluation is inaccurate, and individualized treatment medicine lacks are solved.