158 results about "Vector (molecular biology)" patented technology

The invention belongs to the field of molecular biology and immunology, and particularly relates to an anti-CD3 nano antibody capable of activating the activity of T cells as well as a preparation method and application of the anti-CD3 nano antibody. The invention provides a preparation method of the anti-CD3 nano antibody. The preparation method comprises the following steps: constructing a CD3 plasmid and an overexpression cell line, and immunizing alpaca to obtain peripheral blood mononuclear cells (PBMC) of the alpaca; extracting RNA (Ribonucleic Acid) and carrying out reverse transcription to obtain cDNA; carrying out PCR amplification on the antibody gene segment and cloning the antibody gene segment to a phage display vector; and electrically transforming the vector into SS320 competent cells to obtain the nano antibody library. After two rounds of panning, detecting and screening an antibody clone capable of being combined with the CD3 protein through ELISA (Enzyme-Linked Immunosorbent Assay); an antibody sequence is selected through sequencing, and an antibody protein is expressed by using a mammalian cell high-throughput expression system; and then detecting and screening the antibody which is only specifically combined with the CD3. The method disclosed by the invention is simple, convenient and rapid to operate, and can be used for preparing active antibodies for treating potential tumors and proliferative diseases.

The invention belongs to the technical field of molecular biology and genetics, and particularly relates to a method for editing cattle genes based on Pro-iCHI. The invention provides a method for editing cattle genes based on Pro-iCHI, which comprises the following steps: transiently expressing protamine in cattle AG-haESCs after gene editing, and injecting into mature oocytes to obtain reconstructed embryos. The protamine can remove abnormal DNA methylation generated by an oocyte cytoplasm injection haploid stem cell technology, cell nucleuses are compressed into a sperm-like structure, the obtained bovine Pro-iCHI embryo can smoothly develop into a blastocyst, and the ratio of the blastocyst to the blastocyst obtained by in-vitro fertilization is equivalent. Moreover, the protamine coding gene is inserted into the saccharomyces cerevisiae protein expression vector and then is subjected to transient expression, so that the protamine coding gene cannot be integrated into a genome to cause insertion of an exogenous gene while abnormal DNA methylation is erased.

The invention belongs to the technical field of molecular biology, and particularly relates to a CeNF-YA3 gene, an expression vector and application of the CeNF-YA3 gene in vegetable fat regulation and control. The nucleotide sequence of the CeNF-YA3 gene is as shown in SEQ ID NO.1. After the CeNF-YA3 gene is over-expressed, the grease content of arabidopsis seeds and leaves can be remarkably increased; in addition, the invention also provides a series of vectors containing the CeNF-YA3 gene for subcellular localization, yeast hybridization and plant overexpression for the first time, and proves that the CeNF-YA3 protein has a transcriptional activation function, is localized in a cell nucleus and accords with the basic characteristics of transcription factors for the first time. Therefore, the technical scheme of the invention provides scientific guidance for the application of the CeNF-YA3 gene and the protein thereof.

The invention discloses an AAV vector targeting improvement method applicable to gene therapy drugs, which comprises the following steps: performing high-throughput analysis on target tissue cells through a screening sub-module, screening candidate receptor genes interacting with AAV vector targeting molecules, and determining target receptor genes; the selection submodule is used for designing a gene editing system and selecting a promoter with tissue specific expression to drive the expression of the gene editing system; the introduction submodule is used for introducing the gene editing system into target tissue cells by adopting a gene transfer technology; and the evaluation sub-module is used for selecting the most suitable gene transfer mode according to the characteristics and treatment requirements of the target tissue, and detecting and evaluating the cells subjected to gene editing by utilizing a PCR molecular biological technology. The targeting property of the AAV vector to target tissue cells is improved, the infection of non-target tissues is reduced, and the side effect risk is reduced, so that the gene therapy effect is remarkably improved.

The invention relates to the technical field of molecular biology, in particular to a codon optimization method. The invention provides a codon optimization method, the codon is replaced according to parameters such as codon type and quantity, local GC content, local repetitive sequence, mRNA secondary structure, mRNA free energy and the like, and the optimized sequence can be subjected to experimental screening to obtain a DNA sequence of high-expression protein. The downstream of the method is mainly the application in the IVT mRNA direction: the optimized DNA sequence is constructed into a vector, the plasmid is linearized to serve as a template for in-vitro mRNA transcription, the mRNA is encapsulated into LNP or LNP coupled with other molecules, and the protein expression in 293T cells (not limited to 293T cells) or mice is obviously improved.

The invention relates to the fields of molecular biology and genetic engineering, and relates to a cherax quadricarinatus egg cell guide peptide, a fusion protein, a nucleic acid molecule, a recombinant expression vector, a host cell, a method for guiding a foreign protein to enter a cherax quadricarinatus egg cell and application of the cherax quadricarinatus egg cell guide peptide. The amino acid sequence of the cambarus clarkii egg cell guide peptide is as shown in SEQ ID NO. 1. According to the invention, green fluorescent protein (eGFP) is used as example protein, which proves that the polypeptide can successfully deliver foreign protein. The breakthrough can lay a technical foundation for introducing a gene editing tool (such as a CRISPR-Cas9 system) into a crayfish oocyte, and is expected to be widely applied to the fields of crayfish breeding, functional gene research, aquaculture gene improvement and the like.

The invention discloses a porcine circular RNA, a porcine circLDHC, an overexpression vector and a construction method and application thereof, and relates to the technical field of livestock molecular biology. The porcine circular RNA is formed by cyclizing 3rd to 6th exons of an LDHC gene, and the nucleotide sequence of the porcine circular RNA is as shown in SEQ ID NO: 1. The porcine circLDHC is obtained by carrying out reverse transcription on porcine circular RNA (Ribonucleic Acid). According to the invention, a specific amplification primer is designed for circLDHC, and the real existence of circLDHC is verified through PCR (Polymerase Chain Reaction) amplification, sequencing and RNase R enzyme digestion. Fluorescent quantitative PCR analysis finds that circLDHC is highly expressed in sexual mature testis tissues of pigs, the expression level of circLDHC is related to testis maturation, and meanwhile, the expression quantity of circLDHC in testis of 75-day-old Meishan boars is remarkably higher than that of circLDHC in testis of same-day-old Duroc boars (plt; 0.01) of the substrate. The invention further finds that the over-expressed circLDHC inhibits the proliferation of the porcine sertoli cells, promotes the maturation of the porcine sertoli cells, lays a foundation for further researching the start of the puberty period of the boars, and provides a new thought for analyzing the sexual precocious puberty mechanism of the boars and improving the reproductive performance.

The invention discloses a stropharia rugoso-annulata SRU6 gene promoter, a silent vector containing the promoter and application of silent stropharia rugoso-annulata endogenous genes, and relates to the field of molecular biology and gene engineering. According to the principle of shRNA mediated gene silencing, a silencing vector suitable for gene silencing of stropharia rugoso-annulata is constructed, and meanwhile, a stropharia rugoso-annulata efficient genetic transformation system is established; by utilizing the silent vector and the transformation system established by the invention, the expression quantity of the target gene can be reduced by about 80%, the purpose of gene function research is achieved, and the silent vector and the transformation system have very strong theoretical research and application values.

The invention belongs to the technical field of molecular biology and genetic engineering, and discloses a CRISPR / dCas9 vector for improving the yield of lactyl-N-tetrasaccharide of bacillus subtilis as well as a construction method and application of the CRISPR / dCas9 vector, and a CRISPR / dCas9 system is applied to production of lactyl-N-tetrasaccharide from bacillus subtilis 168 for the first time. According to the invention, pW-dcas9, pW-sgRNA and pW-sgRNA-zwf recombinant vectors are constructed, the transcriptional level of the zwf gene in a phosphopentose pathway is significantly reduced by 11.32 times by using a CRISPR / dCas9 system induced by xylose, and the metabolic flux of a competitive pathway is weakened. The yield of the lactyl-N-tetrasaccharide of the engineering strain obtained by the invention is greatly improved by 31% and reaches 7.25 g / l. Meanwhile, an effective method is provided for lowering metabolic flows of a cell wall competitive pathway and a glycolysis competitive pathway in a later period.

The present invention relates to the field of molecular biology, and in particular to an SLC17A5 gene inhibitor and a use thereof. The present invention provides a use of the SLC17A5 gene inhibitor in preparation of a kidney cancer treatment product. According to the SLC17A5 gene inhibitor provided by the present invention, a proper RNAi target sequence and oligo DNA double-stranded sequence are designed for a target gene, and a lentiviral vector plasmid containing the oligo DNA double-stranded sequence and a finally formed lentivirus are constructed; the resistance gene carried in the lentivirus can effectively reduce the mRNA expression of the target gene SLC17A5, and the lentivirus has an very obvious knockdown effect, has high inhibitory efficiency on cell proliferation and cell migration of kidney cancer cells, and can be used in drugs for treating kidney cancer; moreover, the present invention can be operated with high throughput and has high repeatability.

The invention discloses a methionine nutrition selection marker saccharomyces cerevisiae vector as well as a construction method and application thereof, and relates to the field of genetic engineering and synthetic biology, a methionine MET17 promoter and a coding region sequence are obtained through gene artificial synthesis, a leucine promoter and a coding region sequence in a pAG415-GAL1-ccdB vector are replaced by homologous recombination, and thus the methionine nutrition selection marker saccharomyces cerevisiae vector is obtained. A pAG415-GAL1-MET-ccdB expression vector is constructed and obtained, a ccdB gene is replaced by a green fluorescent protein reporter gene GFP sequence to be integrated into the pAG415-GAL1-MET-ccdB vector, a pAG415-GAL1-GFP-MET vector is constructed and obtained, the obtained expression vector can be used for screening exogenous gene transformants in a methionine deficient culture medium, meanwhile, green fluorescent GFP tag protein carried in the vector has the advantages that the expression vector can be applied to screening of the methionine deficient culture medium, and the expression efficiency of the methionine deficient culture medium is improved. And more application space is provided for carrying out subcellular localization, immunoprecipitation and other downstream inheritance and molecular biology analysis on a target gene in wine brewing cells.

The invention discloses novel application of a GJB5 gene, a construction method of a GJB5 gene overexpression vector and a method for improving high temperature resistance of economic animals, and relates to the technical field of molecular biology. The preparation for overexpressing the GJB5 gene improves the heat resistance of the economic animal sertoli cells through overexpression of the GJB5 gene. Meanwhile, the invention also discloses a construction method of the GJB5 gene overexpression vector and a method for improving the high temperature resistance of economic animals. The effect of the GJB5 gene in heat stress is found for the first time, the gene is expected to be overexpressed in pigs or genetically modified into economic animals such as cattle and sheep in the future, so that the heat resistance of the pigs is improved, the semen quality of the pigs under the heat stress condition is further improved, and the GJB5 gene has important social benefits and economic benefits.

The invention relates to a recombinant expression vector of a human antibacterial peptide, an engineering bacterium, a construction method and application, and relates to the technical field of molecular biology. The structure of the recombinant expression vector is a pET28a-sfGFP-HRV3C tangency point-LL37, and the nucleotide sequence of the LL37 in the pET28a-sfGFP-HRV3Cp tangency point-LL37 is as shown in SED ID NO: 1, and the nucleotide sequence of the LL37 in the pET28a-sfGFP-HRV3Cp tangency point-LL37 is as shown in SED ID NO: 2. And the sfGFP in the pET28a-sfGFP-HRV3Cp tangency point-LL37 is the super-folded green fluorescent protein. The invention further discloses a preparation method of the super-folded green fluorescent protein. Compared with the prior art, an expression system with a fusion tag is designed by combining a fusion expression strategy and protein characteristics of LL37, so that soluble expression of LL37 in escherichia coli is promoted, and efficient production of target protein is realized; compared with an existing target protein obtaining mode, the method has the advantages that the protein expression quantity is increased by using sfGFP, and the method is more suitable for large-scale industrial application.

This application relates to the field of molecular biology, and more specifically to isolated transposases and their use. More specifically to nucleic acids and nucleic acid constructs encoding transposases, nucleic acid sets and nucleic acid set constructs, and compositions, recombinant vectors, recombinant host cells and kits comprising transposases. More specifically to methods for introducing exogenous nucleic acid fragments into the genome of host cells, methods for editing the genome of host cells, and methods for obtaining host cells containing exogenous nucleic acid fragments in their genomes. More specifically to the use of transposases, nucleic acids and nucleic acid constructs, nucleic acid sets and nucleic acid set constructs, compositions, recombinant vectors, or recombinant host cells for introducing exogenous nucleic acid fragment genes into the genome of host cells, or for preparing drugs or formulations for gene therapy, cell therapy, genome research, or stem cell induction and post-induction differentiation.

The invention relates to a Sebastes schlegeli endogenous retrovirus envelope protein Penv, a lentiviral vector and application thereof, and belongs to the field of genetic breeding of molecular biology, and the amino acid sequence of the envelope protein Penv is as shown in SEQ ID NO.1. The invention further provides an in-vitro assembly system, a transformant and a kit containing membrane fusion protein particles and lentiviral particles of the envelope protein Penv and application of the envelope protein Penv. VSVG protein is replaced with Penv protein from sclerobone fish, so that the transduction efficiency of the lentiviral particles to sclerobone fish cells is effectively improved, and the transduction efficiency of the lentiviral particles to the sclerobone fish cells is improved. And efficient gene delivery is realized. Meanwhile, as the endogenous Env protein, the Env protein overcomes the immunological rejection of a host to the exogenous Env protein, and also has the potential of in-vivo application.

The invention belongs to the technical field of intelligent prediction, and relates to a key gene identification method and device, computer equipment and a storage medium, and the method comprises the steps: obtaining original sample data; preprocessing the original sample data to obtain preprocessed sample data; performing word segmentation operation on the preprocessed sample data according to the HFkme r to obtain word segmentation data; performing vector conversion operation on the word segmentation data to obtain word segmentation vector data; constructing an initial BGAC model, and performing model training operation on the initial BGAC model according to the word segmentation vector data to obtain a target BGAC model; obtaining a to-be-recognized genome protein sequence; and inputting the genome protein sequence into the target BGAC model for key gene recognition operation to obtain a key gene recognition result. The method not only solves the problems of high time and high cost in key gene identification in traditional molecular biology, but also provides a precious microscopic view angle for deeply exploring the origin and evolution of insect wings, and the complex biological macro-evolution process is better understood.

The invention belongs to the technical field of molecular biology, and particularly relates to a method for transferring an exogenous nucleic acid fragment into a plant by using a plant virus. The technical problem to be solved by the invention is to provide a new method which can be applied to transferring an exogenous fragment into a plant. According to the technical scheme, the method for transferring the exogenous nucleic acid fragment into the plant by using the plant virus comprises the following steps: fusing the exogenous nucleic acid fragment into a 3'non-coding RNA region of TVMVwt to obtain a fused fragment, constructing the fused fragment onto an infectious clone skeleton vector, and transforming the plant. According to the invention, an exogenous coding sequence is inserted into a 3'non-coding RNA region of TVMVwt to construct an infectious cloning vector, the infectious cloning vector is transferred into a plant, and expression of the exogenous coding sequence is realized.

The present application relates to the field of molecular biology, in particular to a recombinant human heparin N-deacetylase encoding gene, a recombinant vector, a recombinant strain and application thereof, the recombinant human heparin N-deacetylase constructed in the expression process shows excellent stability and activity, has the ability of specific catalysis of deacetylation of acetyl group in heparin precursor, and can effectively generate deacetylated heparin precursor. This property significantly improves the conversion efficiency and reduces the generation of by-products. The present application optimizes the codon of the recombinant human heparin N-deacetylase, and uses the promoter P AOX1 or P GAP to regulate the expression, so that the expression level of the recombinant human heparin N-deacetylase in Pichia pastoris is significantly improved, and the obtained GS115-AOX1-hNDase and GS115-PGAP-hNDase strains successfully realize the secretory expression of the target gene, providing a new strategy and method for the biosynthesis of heparin.

This application belongs to the field of plant molecular biology technology and specifically provides a PsbHLH35 gene and encoded protein, and their use in tree peony somatic embryogenesis. A PsbHLH35 gene, comprising the nucleotide sequence shown in SEQ ID NO. 1. A protein encoded by the PsbHLH35 gene, comprising the amino acid sequence shown in SEQ ID NO. 2. A method for using the PsbHLH35 gene in tree peony somatic embryogenesis, comprising the following steps: S1) cloning the PsbHLH35 gene into a vector to construct a PsbHLH35 gene overexpression vector; S2) transforming the PsbHLH35 gene overexpression vector into tree peony. The PsbHLH35 gene of this application has a significant promoting effect on tree peony somatic embryogenesis.

The present invention belongs to the field of molecular biology and genetics, and in particular relates to a method for editing a bovine gene based on Pro-iCHI. The present invention provides a method for editing a bovine gene based on Pro-iCHI. Protamine is transiently expressed in gene-edited b-haSCs, which are then injected to mature oocytes to obtain reconstructed embryos. Protamine can eliminate abnormal DNA methylation resulting from oocyte intracytoplasmic haSCs injection and enable the nucli to compress into sperm-like structures, and the obtained bovine Pro-iCHI embryos can successfully develop into blastocysts, with a blastocyst rate comparable to that of the embryos obtained by in vitro fertilization. Moreover, in the present invention, a protamine-encoding gene is inserted into a Saccharomyces cerevisiae protein expression vector for transient expression, which ensures that abnormal DNA methylation is erased, without integration into the genome resulting in the insertion of exogenous genes.

This invention discloses a novel site-specific recombinase-mediated multisite chromosome editing tool and its applications. The invention provides a complete product suite for integrating or continuously integrating the genome of a host cell, comprising: a circular vector 1 composed of the following elements: loxF1, nox, ori, resistance gene, lox71, P3, P1, P2; and / or a circular vector 2 composed of the following elements: lox66, nox, ori, resistance gene, loxF2, P3, P1, P2; and a circular vector 3 for expressing Nigri and Cre recombinases; and a site-specific knock-in tool for knocking in the lox66-nox fragment at the desired integration site in the host cell genome; P1 and P2 are adapters, used to insert the DNA fragment to be integrated, and P3 is a universal primer. This invention enables rapid and continuous genome integration using simple molecular biology operations.

The invention is suitable for the technical field of molecular biology, and provides an SlSYP121 gene and application of a protein coded by the SlSYP121 gene in tomato leaf shape development. The nucleotide sequence of the SlSYP121 gene is as shown in SEQ ID No.1 (Sequence Identifier Number 1). The amino acid sequence of the SYP121 protein coded by the SlSYP121 gene is as shown in SEQ ID No. 2. According to the invention, the SlSYP121 overexpression vector is successfully introduced into a plant body, and a positive overexpression plant is screened out. Compared with a wild type plant, the positive overexpression plant has a leaf growth and development form different from that of the wild type plant, the phenomena that the leaf notch degree is reduced, and the epicotyl length and the leaf mucoid glandular hair length are reduced occur, and the morphological change lays a foundation for subsequent overexpression plant stress resistance experiments.

The invention relates to the technical field of crossing of molecular biology, nucleic acid detection and bioinformatics, and discloses an in-vivo detection method for synchronously detecting CRISPR off-target sites and viral vector integration sites. According to the in-vivo detection method for synchronously detecting the off-target site of the CRISPR and the integration site of the virus vector, the reverse terminal repeat (ITR) of the AAV vector is used as a tag sequence for the first time, and a universal PCR (Polymerase Chain Reaction) amplification primer is designed and a unique molecular identifier (UMI) technology is introduced to detect the off-target site of the CRISPR and the integration site of the virus vector. And meanwhile, an off-target event and an AAV carrier integration event in an in-vivo CRISPR editing process are captured and accurately recognized. Compared with the existing schemes such as GUIDE-seq, DISCOVER-Seq +, OliTag-seq, U-DiTaS and the like, the in-vivo detection method for synchronously detecting the CRISPR off-target site and the virus vector integration site, provided by the invention, has the advantage that remarkable breakthrough is realized in the aspects of in-vivo applicability, detection depth, operation economy and industrialization standardization.

The invention belongs to the technical field of molecular biology, and particularly relates to an anti-GLP-1R nano antibody as well as a preparation method and application thereof. The invention provides an anti-GLP-1R nano antibody, and also provides a construction method of the anti-GLP-1R nano antibody. The method comprises the following steps: constructing GLP-1R overexpression plasmids and overexpression cell strains, immunizing alpaca by using the plasmids and the cells to obtain PBMC cells, extracting total RNA, carrying out reverse transcription to obtain cDNA, amplifying antibody gene segments through PCR, connecting the segments to phage expression vectors, and electrically transforming the constructed vectors into SS320 competent cells to finally obtain the GLP-1R immune antibody library. The method comprises the following steps: carrying out cell panning for one round, carrying out Cell ELISA detection to obtain an antibody combined with GLP-1R, carrying out sequencing to obtain an antibody sequence, obtaining antibody protein through mammalian cells, and carrying out FACS detection by using CHOK1-GLP-1R cells to finally obtain the antibody combined with GLP-1R. The method is simple and convenient to operate, and the activity of the antibody obtained by using a mammalian cell expression system is closer to that of a natural antibody.

The present invention relates to the technical field of molecular biology, and in particular, to an anti-CD40 nanobody, and a preparation method therefor and use thereof. The anti-CD40 nanobody provided by the present invention has three unique complementarity determining regions, namely CDR1, CDR2, and CDR3. The present invention further provides a sequence encoding the nanobody or its VHH chain, a corresponding expression vector, and a host cell capable of expressing the nanobody. The present invention includes immunizing an alpaca by constructing a CD40 antigen to obtain a PBMC of the alpaca and constructing a vector; screening an anti-CD40 antibody through phage display; and finally obtaining a highly sensitive and specifically binding antibody through functional assays and sequencing result analysis. The nanobody provided by the present invention can specifically recognize and bind to CD40.

The present invention relates to the fields of molecular biology and genetic engineering, and relates to crayfish egg cell guide peptides, fusion proteins, nucleic acid molecules, recombinant expression vectors, host cells, methods for guiding exogenous proteins into crayfish eggs, and applications thereof. The amino acid sequence of the crayfish egg cell guide peptide is as shown in SEQ ID NO.1. The present invention uses green fluorescent protein (eGFP) as an example protein to prove that the polypeptide can successfully deliver exogenous proteins. This breakthrough can lay a technical foundation for the introduction of gene editing tools (such as the CRISPR-Cas9 system) into crayfish oocytes, and is expected to be widely used in the fields of crayfish breeding, functional gene research, and aquaculture gene improvement.

The present invention discloses circApbb1 molecules, siRNA targeting circApbb1 expression, adeno-associated virus vectors, preparation methods and applications, belonging to the field of molecular biology applications. A siRNA capable of knocking down circApbb1 expression was designed, and an adeno-associated virus vector HBAAV2 / 9‑cTNT‑mir30‑circApbb1Green of the siRNA was constructed. Detection showed that circApbb1 was significantly upregulated in the hearts of mice with atherosclerosis. The prepared siRNA adeno-associated virus vector can effectively reduce the expression of circApbb1 in the mouse heart, reduce the area of ​​atherosclerotic plaques, inhibit the progression of atherosclerosis and improve cardiac function, thus realizing the use of circApbb1 to treat atherosclerosis, providing an important supplement to the biological function of circApbb1, and providing a new technical means for the treatment of atherosclerosis.

The invention belongs to the technical field of molecular biology, and particularly relates to a recombinant vector for expressing OrfV truncated F1L protein, a strain and application of the recombinant vector. The preparation method comprises the following steps: truncating an F1L gene of an orf virus, introducing a His tag and a termination codon sequence, and constructing into a pYD1 plasmid to obtain a pYD1-F1L recombinant plasmid; and introducing the recombinant plasmid into competent cells of saccharomyces cerevisiae to obtain the recombinant yeast strain. According to the invention, the F1L gene of OrfV is truncated, so that the immunogenicity of the OrfV is effectively retained; the anchoring protein is displayed on the surface of the saccharomyces cerevisiae, so that the immune function of the saccharomyces cerevisiae can be developed; recombinant saccharomyces cerevisiae is adopted as a delivery carrier, the antigen protein can better play an immune role by means of the constant value of the recombinant saccharomyces cerevisiae in intestinal tracts, and meanwhile, the risk that the F1L protein is degraded by protease when being orally taken is further reduced. The invention provides a new direction for the development of OrfV oral vaccines.

The invention discloses a ribosome group or selective ribosome group detection and analysis method based on an immune principle, and relates to the field of molecular biology and genomics. Comprising the following steps: sample preparation and cross-linking: treating cells by using a translation inhibitor to suspend ribosome translation, and then cross-linking RNA-protein in the cells by using a membrane permeable cross-linking agent; then cracking the cells, digesting RNA of a cracking product by using ribonuclease, and retaining ribosome and mRNA fragments protected by the ribosome; immune enrichment: carrying out specific immune enrichment on the digested sample by using an antibody combined with a carrier, and eluting to obtain a ribosome-mRNA compound; then decrosslinking and removing protein, extracting mRNA fragments and carrying out phosphorylation treatment so as to add joints, build a library and carry out sequencing; and finally, library construction, sequencing and bioinformatics analysis are carried out. The method has the advantages of high-specificity enrichment, accurate positioning of translation sites, wide application range and accurate sequence information.

The invention aims to provide a co-expression recombinant vector, a recombinant strain and a method for enzymatic synthesis of 6-selenium guanine nucleotide, and belongs to the technical field of molecular biology. The method provided by the invention has the advantages of low cost, specificity, simplicity, rapidness, low reaction condition requirements, no environmental pollution and the like. According to the co-expression recombinant vector provided by the invention, the soluble expression effect of YcfA or YcfAD166R / Y170N protein and YcfC protein can be greatly improved, a further constructed recombinant strain can be used for developing engineering strains for synthesizing compounds such as 6Se-GTP, 6Se-GDP, 6Se-GMP and the like in bacteria, and the co-expression recombinant vector has a wide application prospect.