864 results about "Point mutation" patented technology

The invention provides a carbonyl reductase mutant. The carbonyl reductase mutant is obtained by performing single-point mutation or multi-point combined mutation on the 17th site, the 40th site and the 64th site of an amino acid sequence as shown in SEQ ID NO.1. The invention also provides a coding gene, a recombinant vector containing the coding gene, a co-expression engineering bacterium and an application of the co-expression engineering bacterium. Compared with a wild type enzyme, the carbonyl reductase mutant has high activity and high stereoselectivity, the enzyme activity can reach more than two times that of the wild type enzyme, the catalytic efficiency on a substrate precursor ketone is remarkably improved, the yield of a statin drug intermediate and chiral alcohol synthesized by an enzyme method of the carbonyl reductase mutant is remarkably improved, and the yield of a rosuvastatin intermediate (3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R, 3R) is remarkably improved. The highest yields of the (3R, 5R)-6-chloro-3, 5-dihydroxyhexanoic acid tert-butyl ester and the atorvastatin intermediate (3R, 5R)-6-cyano-3, 5-dihydroxyhexanoic acid tert-butyl ester reach 99% and 97.6% respectively, e.e. Is larger than 99%, and the method has high industrial application value.

The invention discloses polyketide synthase or a polyketide synthase mutant and application thereof, and belongs to the technical field of gene engineering. The polyketide synthase is AspiPKS4, AspiPKS5, AspiPKS6 and AspiPKS7 from cyathea spinulosa, FhPKS2, FhPKS3, FhPKS4, FhPKS5 and FhPKS6 from ficus himalayana, and CrPKS from floating fern. The mutant is obtained by carrying out single point mutation on the second amino acid of AspiPKS7 or deleting an N-terminal redundant sequence. The polyketide synthase or the mutant is used for replacing NnHisspS in an original FBP luminescence system, and the biological self-luminescence level can be remarkably improved. The invention further provides a simplified and improved FBP system which comprises the polyketide synthase or the polyketide synthase mutant, CPH, H3H and Luz, and the application convenience and the application luminescence capacity of the FBP system are improved.

The invention discloses a glycosyl transferase mutant and an application of the glycosyl transferase mutant in synthesis of rebaudioside. According to the invention, single-point mutation and multi-point mutation are carried out on the basis of a glycosyl transferase amino acid sequence as shown in SEQ ID NO: 1, and a mutant with improved catalytic activity is obtained. The catalytic activity, the substrate specificity and / or the substrate specificity of the glycosyl transferase mutant are / is changed, and the catalytic activity of enzyme to a specific substrate can be remarkably improved by mutation at a specific site. And carrying out induced expression and protein purification on the obtained mutation sequence to obtain the mutant enzyme. The mutant enzyme is used as a catalyst, and UDPG is used as a glycosyl donor, so that the catalytic reaction efficiency of substrates such as stevioside ST, rebaudioside A (RebA) and rebaudioside D (RebD) can be obviously improved. According to the glycosyl transferase UGT76G1 mutant constructed by the research, the catalytic activity of the glycosyl transferase UGT76G1 mutant is improved, and efficient production of rebaudioside M is realized by optimizing a reaction system.

The invention discloses a hemoglobin-resistant Taq DNA polymerase mutant and a construction method thereof, and relates to the field of biology, and the hemoglobin-resistant Taq DNA polymerase mutant is characterized in that a nucleotide sequence for coding the Taq DNA polymerase is shown as SEQ ID NO.1, and the mutant comprises at least one mutation or all mutations selected from S623D and E721A sites. On the basis of natural Taq DNA polymerase, the molecular structure of the Taq DNA polymerase is modified through rational design and combination with a site-specific mutagenesis biotechnology, so that the Taq DNA polymerase mutant is more suitable for PCR amplification of samples containing hemoglobin and the like than a wild type, and the result judgment accuracy of a sample containing a hemoglobin inhibitor is more facilitated.

The invention discloses a thermostable conjugated polyketoreductase mutant, which is obtained by performing single-point mutation or multi-point combined mutation on the 25th site, the 92th site, the 214th site or the 256th site of an amino acid sequence as shown in SEQ ID NO.1. The invention further discloses a preparation method of the thermostable conjugated polyketoreductase mutant. The initial enzyme activity of the mutant is high, and the residual enzyme activity is gt after incubation at 50 DEG C; the Tm value can be increased by more than 9 DEG C compared with that of an original strain, the 50 DEG C half-life period t1 / 2 is increased by more than 377.8 times, and the thermal stability is obviously improved. The invention also provides a coding gene, a recombinant vector containing the coding gene and a co-expression engineering bacterium. The thermostable conjugated polyketoreductase mutant, glucose dehydrogenase and L-pantolactone dehydrogenase are used for constructing a three-enzyme co-expression engineering bacterium as a biocatalyst, racemization pantolactone is catalyzed to synthesize DPL, for a 1M substrate, the yield of the DPL reaches 97.5% within 36 hours, KPL hydrolysis is avoided, the reaction time is shorter, the product concentration is higher, and the thermostable conjugated polyketoreductase mutant has great market application value.

The invention relates to the technical field of gene engineering and enzyme engineering, in particular to an esterase mutant for degrading ethyl carbamate and application of the esterase mutant, the esterase mutant has an amino acid sequence as shown in SEQ ID NO.1 which is subjected to site mutation, the site mutation is any one or more site combined mutation of V129S, V161L or I229M, and the amino acid sequence of the esterase mutant is shown in SEQ ID NO.1. The V129S site mutation is that valine at the 129 site is mutated into serine, the V161L site mutation is that valine at the 161 site is mutated into leucine, and the I229M site mutation is that isoleucine at the 229 site is mutated into methionine. Mutation is carried out through a site-directed mutagenesis method, so that the amino acid sequence of the esterase mutant V129S / I229M is changed, the autoplasmic structure and function of eggs are changed, the esterase mutant V129S / I229M with the ethyl carbamate hydrolysis rate increased to 6.55 times in an acid environment is finally obtained, and the esterase mutant V129S / I229M has high industrial application value.

The invention provides a molecular marker related to immune traits of elephant male semi-fine wool sheep and a detection primer and application thereof, and belongs to the technical field of molecular assisted breeding. The nucleotide sequence of the molecular marker is shown as SEQ ID NO.1; the molecular marker comprises an SNP site, the SNP site is located at the 117th site of the molecular marker, the mutation basic group of the SNP site is A or G, and the immune globulin content of an individual male semi-fine wool sheep with the genotype of AG or GG at the SNP site is remarkably higher than that of an individual with the genotype of AA. The content of immune globulins IgA, IgG and IgM of an individual elephant male semi-fine-wool sheep can be judged according to the genotype of the molecular marker, the immune traits of the elephant male semi-fine-wool sheep can be rapidly and accurately evaluated, the high-immunity elephant male semi-fine-wool sheep can be screened, the molecular marker has important significance on early-stage auxiliary breeding of the elephant male semi-fine-wool sheep, the breeding period is remarkably shortened, and the breeding cost is reduced. The breeding efficiency is improved.

The invention discloses a high-efficiency lysine decarboxylase mutant and an application of the high-efficiency lysine decarboxylase mutant in synthesis of pentamethylene diamine. A series of mutants including a single mutant (such as N218G, T222V, C244P and the like), a double mutant and a three mutant are obtained by carrying out multi-site mutation on wild type lysine decarboxylase (SEQ ID NO.2) from Klebsiella grimontii. The catalytic efficiency of the mutant is remarkably improved, and the activity of the mutant is improved by 5.0-10.2 times compared with that of a wild type enzyme. According to the method disclosed by the invention, the substrate conversion rate of 99% or above can be realized within 2 hours through an optimized process of constructing an expression vector and a genetically engineered bacterium, inducing expression of lysine decarboxylase and synthesizing 1, 5-pentanediamine by utilizing a whole-cell catalysis technology. Compared with a traditional chemical method and a natural enzyme method, the method disclosed by the invention has the advantages of simplified process, high catalytic efficiency, low production cost and the like, and is suitable for producing bio-based 1, 5-pentamethylene diamine and nylon 56 salt.

The invention discloses a grass carp gamma interferon fusion protein as well as a mutant, a coding gene and application thereof. In order to improve the expression quantity or expression efficiency of the grass carp gamma interferon, codon optimization and mutation are carried out on the coding gene of the grass carp gamma interferon to obtain the grass carp gamma interferon mutant. The fusion protein is further obtained by connecting the grass carp gamma interferon mutant with the C end of the grass carp ferritin heavy chain subunit of which the last 17 amino acids are removed through a connecting peptide. In order to further improve the antiviral activity of the fusion protein, the obtained fusion protein is subjected to single-point mutation, and the titer of the fusion protein is remarkably improved. The fusion protein is expressed by a silkworm cell eukaryotic expression system, and the fusion protein shows a conformation suitable for the interferon to play functions based on the ferritin self-assembly characteristic, so that the titer of the interferon in a host is effectively improved, and the in-vivo and in-vitro half-life period of the interferon is prolonged. The invention has application prospects in preparation of drugs or reagents for preventing or treating viral diseases of grass carp and the like.

The present disclosure relates in some aspects to methods for analyzing target nucleic acids and their spatial locations in a biological sample. In some aspects, the presence / absence, amount, and / or identity of variant sequences (e.g., single nucleotide variations such as SNPs or point mutations) in a plurality of target nucleic acids in a cell or tissue sample are analyzed in situ in the sample. Also provided are compositions and kits for use in accordance with the methods.

The invention belongs to the technical field of microbial genetic engineering, and particularly relates to an alpha-2, 3-sialyltransferase mutant and application thereof. The a-2, 3-sialyltransferase mutants V175N, P198A, G173D, D174Q, F196K and A315H are obtained by performing site-specific mutagenesis on six amino acids in an a-2, 3-sialyltransferase amino acid sequence coded by a gene derived from Campylobacter jejuni (OH4384), the yield of 3 '-sialyllactose is remarkably increased by single-point mutagenesis of the V175N, the yield of 3'-sialyllactose is remarkably increased by single-point mutagenesis of the V175N, the yield of 3 '-sialyllactose is remarkably increased by single-point mutagenesis of the P198A, the G173D, the D174Q, the F196K and the A315H, the optimal recombinant microbial strain 3 '-SL-09 (V175N / P198A / D174Q) is obtained through combination of multiple mutation sites, the yield can reach 5.91 g / L, the bottleneck problem of low catalytic efficiency of sialyltransferase is effectively solved, and an enzyme element with excellent performance is provided for high-efficiency and low-cost biological manufacturing of 3'-SL.

The invention discloses a 3 '-phosphoadenosine-5'-phosphoryl sulfate synthetase mutant and an application of the 3 '-phosphoadenosine-5'-phosphoryl sulfate synthetase mutant. Compared with a wild enzyme amino acid sequence as shown in SEQ ID NO.1, the amino acid sequence of the 3 '-adenosine-5'-phosphoryl sulfate synthetase mutant has one or two mutations in the 101 site, the 207 site and the 560 site; preferably, phenylalanine at the 101 site is mutated into tryptophan; cysteine at the 207th site is mutated into glycine; phenylalanine at the 560th site is mutated into tryptophan. The invention also provides a synthesis method and application of the 3 '-adenosine-5'-phosphoryl sulfuric acid. The APS intermediate product inhibition effect of hPAPSS1 is relieved through a point mutation technology, the substrate affinity of hPAPSS1 to ATP is improved, and a mutant enzyme with a high PAPS conversion rate is obtained; and the bifunctional enzyme hPAPSS1 is synthesized by heterologous expression of PAPS in escherichia coli, so that the acquisition of the enzyme is simplified, and the synthesis efficiency of PAPS is improved.

The present invention provides an ATP phosphoribosyltransferase mutant and its application in histidine production. The mutant is obtained by performing point mutations at positions 84 and / or 205 on the amino acid sequence of the wild-type ATP phosphoribosyltransferase derived from Acinetobacter baumannii. The mutant, the nucleic acid molecule encoding the mutant, the expression cassette containing the nucleic acid molecule, the recombinant vector or the recombinant microorganism are used for the production of L-histidine, and the yield of L-histidine is significantly increased, laying a good foundation for the large-scale industrial production of L-histidine.

The present invention provides a 4-vinylguaiacol oxygenase mutant for synthesizing vanillin, and an engineered bacterium thereof. The 4-vinylguaiacol oxygenase mutant is an enzyme having the sequence as shown in SEQ ID NO. 1, which exhibits a double-site mutation of D27P / T216M, D27P / M351F, D27P / Y156D, M351F / Y156D, T216M / Y156D or T216M / M351F, or a single-site mutation of T216M, T216L, S280M, G307P or M351F, and can be used for catalyzing 4-vinylguaiacol to generate vanillin.

The invention belongs to the technical field of biological pharmacy, and particularly relates to influenza virus-like particles for chimeric expression of new coronavirus RBD protein as well as a preparation method and application of the influenza virus-like particles. The virus-like particle is prepared by embedding fusion protein on the surface of influenza virus M1, and the fusion protein comprises an HA2 segment of HA protein of influenza virus, an extracellular functional region of M2e matrix protein, a conserved region short peptide of nucleoprotein, and RBD tandem protein with mutated five sites of 417, 452, 478, 484 and 501 of new coronavirus; the virus-like particles can be used for preventing specific influenza and new crowns; the preparation method of the virus-like particles is simple and low in cost, the virus-like particles are good in immunogenicity and cross protectiveness, the preparation requirements of large-scale influenza vaccines can be met, the biological safety is high, and the immunity of an organism to various influenza viruses and new coronal viruses can be stimulated.

Disclosed is a Taq DNA polymerase mutant. Compared to a wild-type Taq DNA polymerase, the Taq DNA polymerase mutant has a mutation at one or more of amino acid positions 116, 439, and 468, wherein the amino acid sequence of the wild-type Taq DNA polymerase is set forth in SEQ ID NO: 1. Compared to the wild-type Taq DNA polymerase, the Taq DNA polymerase mutant of the present invention has higher enzyme activity than the wild-type enzyme; both single-site mutants and multi-site mutants show improved tolerance to corn leaf supernatant, mouse serum, and / or heparin sodium, and thus can be used for direct rapid amplification of samples.

The invention belongs to the technical field of bioengineering, and particularly relates to a polyphosphate kinase mutant and application thereof in synthesis of beta-nicotinamide mononucleotide, and the polyphosphate kinase mutant is obtained by single-point or multi-point mutation of 99th, 186th, 190th and 204th amino acids of polyphosphatase with an amino acid sequence shown as SEQ ID NO.1. The invention also relates to a preparation method of the polyphosphate kinase mutant, and the application of the polyphosphate kinase mutant in synthesis of beta-nicotinamide mononucleotide in synthesis of beta-nicotinamide mononucleotide in synthesis of beta-nicotinamide mononucleotide in synthesis of beta-nicotinamide mononucleotide. Polyphosphokinase derived from Lampropedia Hyalina DSM 16112 is subjected to molecular modification to obtain multiple mutants, the enzyme activity of the mutants is improved by 1.5-14 times compared with that of wild type polyphosphokinase, the preference of the mutants to AMP is improved by 1.08-4.88 times compared with that of the wild type polyphosphokinase, and an ATP regeneration system constructed by the mutants can remarkably reduce the use amount of enzyme and reduce the fermentation capacity and cost; the reaction time can be greatly shortened, the influence of AMP on the enzyme activity is reduced, the requirements of large-scale industrial production of NMN prepared by adopting a biological enzyme method can be met, and the method has a wide application prospect.

The invention discloses a dihydrodaidzein reductase mutant and application of the dihydrodaidzein reductase mutant in synthesis of (3S, 4R)-tetrahydrodaidzein, and belongs to the technical field of biology. According to the mutant, wild-type dihydrodaidzein reductase DHDR is taken as a parent, single-point mutation or combined mutation is carried out, and a DHDR mutant is obtained; the parent site of the single point mutation is one of the 120th site, the 122nd site, the 170th site, the 178th site, the 214th site, the 219th site, the 220th site or the 220th site; the combined mutation adopts two modes: one mode is that a single-point mutation DHDR mutant is taken as a parent, mutation is carried out, and a combined mutation mutant I is obtained; and 2, taking the DHDR combined mutation mutant I as a parent, and performing mutation to obtain a combined mutation mutant II. The DHDR mutant disclosed by the invention is used for synthesizing (3S, 4R)-tetrahydrodaidzein by reducing dihydrodaidzein, and asymmetric reduction with high activity and high stereoselectivity can be realized.

The invention belongs to the technical field of genetic engineering, and particularly relates to a method for creating a corn dwarfing material and application of the corn dwarfing material. The invention discloses a method for creating a corn dwarfing material, which specifically comprises the following steps: carrying out specific key site editing on a target gene ZmGA20ox3 and / or ZmGA20ox5 in corn by using a genetic engineering means, and carrying out specific amino acid site mutation on a protein coded by the target gene ZmGA20ox3 and / or ZmGA20ox5 subjected to specific key site editing, and further cultivating and screening to obtain the corn dwarfing material. The method provided by the invention can be used for cultivating transgenic corn plants with different plant heights and other agronomic traits, and meanwhile, a new breeding technical scheme is provided for creating dwarf corn, reducing the height of a corn ear, reducing the included angle of corn plant leaves and increasing the corn yield; crops can be more densely planted in agriculture, so that the yield of the crops in unit area is improved, and the method has important breeding significance.

The invention provides p-hydroxymandelic acid synthetase, a gene, a mutant of the p-hydroxymandelic acid synthetase and application of the p-hydroxymandelic acid synthetase in asymmetric synthesis of (S)-mandelic acid. The p-hydroxymandelic acid synthetase disclosed by the invention is derived from actinomycetes Actinokineospora auranticola, the amino acid sequence of the p-hydroxymandelic acid synthetase is as shown in SEQ ID NO.1, and a mutant of the p-hydroxymandelic acid synthetase is obtained by performing single-point mutation or multi-point combined mutation on glutamic acid at the 187th site, isoleucine at the 189th site and threonine at the 211th site of the amino acid sequence as shown in SEQ ID NO.1. The p-hydroxymandelic acid synthetase and the mutant thereof can convert phenylpyruvic acid into (S)-mandelic acid with high optical purity, have the advantages of mild reaction conditions, simplicity and convenience in operation, safety, environment friendliness and the like, and have a very good industrial application prospect.

The invention provides a nucleotide mutation site prediction model construction and disease-related point mutation identification method. Specifically, the invention provides a deep learning model-fused nucleotide mutation site prediction model construction method and a disease-related point mutation identification method. According to the method, DNA point mutation and RNA point mutation can be recognized from transcriptome sequencing data in a high-sensitivity and high-specificity mode, and basic data is provided for explaining mutation generation mechanisms and functions on the whole transcriptome and genome level.

The invention discloses a VpCas9 protein double-site mutant and an application of the VpCas9 protein double-site mutant in gene editing. The invention firstly provides a homologue VpCas9 protein of Cas9, the amino acid sequence of the homologue VpCas9 protein is shown as SEQ ID No.5, and compared with SpCas9, the homologue VpCas9 protein has a better editing effect in the aspects of gene editing efficiency, PAM compatibility or specificity and the like; in order to further improve the gene editing effect of the VpCas9 protein in genome editing, the VpCas9 protein is subjected to double-site mutation, the VpCas9 protein double-site mutant is obtained, and compared with SpCas9 protein or VpCas9 protein, the VpCas9 protein double-site mutant has a better gene editing effect in genome editing, and the gene editing effect of the VpCas9 protein is improved. The method has an important application prospect in genome editing of plants, animals or microorganisms.

The invention discloses a method for biosynthesizing epigallocatechin gallate glucoside, and belongs to the technical field of biocatalytic synthesis. According to the method, a sucrose phosphorylase mutant is adopted as a catalyst, the sucrose phosphorylase mutant is subjected to single-point mutation or combined mutation on the basis of an amino acid sequence SEQ ID NO.1, and the enzyme activity of the sucrose phosphorylase is improved through mutation; the mutant is utilized to realize efficient catalytic synthesis of epigallocatechin gallate glucoside, and in the whole catalytic process, the synthesis method is mild in condition, simple to operate, short in reaction time and high in catalytic efficiency, and has a good application prospect.

An engineered bacterium for improving tryptophan yield, a construction method therefor, and a use thereof. A strain capable of tolerating a high concentration of tryptophan was obtained by screening. Genome sequencing and protein sequence analysis of the strain revealed that some proteins in the strain had undergone point mutations, and these mutants can improve the tryptophan yield. In order to further improve the tryptophan yield, modifications are made to a protein sequence expressed by a fadR gene or a protein sequence expressed by a pepD gene in a starting bacterium. Such modifications enable the obtained engineered bacterium to have a higher tryptophan yield than the starting bacterium. In the case of large-scale production, in a 5 L fermentation tank, the tryptophan yield can reach 62.38±5.80 g / L, and the sugar-to-acid conversion rate can reach 24.1%. Compared with the original bacterium, the tryptophan yield is increased by 1.48 folds, and the sugar-to-acid conversion rate is increased by 1.26 folds. The biological material and the use thereof relate to the technical field of molecular biology and have wide practical application value.

The invention belongs to the technical field of biosynthesis, and particularly relates to an engineering bacterium for preparing stevioside through de novo fermentation as well as a preparation method and application of the engineering bacterium. According to the invention, mevalonate kinase in the MVA pathway is mutated and optimized. The method comprises the following steps: carrying out point mutation on mevalonate kinase MvK of a wild type source, constructing a stevioside synthesis route in an escherichia coli host, and comparing the influence of MvK mutation on the yield of stevioside, so as to determine a Q160L mutant, and applying the Q160L mutant to the stevioside production route to realize efficient production of stevioside.

The present disclosure relates in some aspects to methods for analyzing target nucleic acids and their spatial locations in a biological sample. In some aspects, the presence / absence, amount, and / or identity of variant sequences (e.g., single nucleotide variations such as SNPs or point mutations) in a plurality of target nucleic acids in a cell or tissue sample are analyzed in situ in the sample or using a spatial array. Also provided are oligonucleotides, sets of oligonucleotides, compositions, and kits for use in accordance with the methods.

The invention discloses a lytic polysaccharide monooxygenase mutant as well as a preparation method and application thereof. In order to solve the problems of low expression level, poor stability and the like of the existing lysing polysaccharide monooxygenase in a microbial expression system, the lysing polysaccharide monooxygenase RsLPMO9 is mutated to obtain a multi-site mutant of which the amino acid sequence is shown as SEQ ID NO.2, and the expression quantity of the multi-site mutant is obviously higher than that of a wild type protein RsLPMO9. The analysis of the thermal stability shows that the thermal stability of the multi-site mutant is obviously improved compared with that of the wild protein RsLPMO9, and the lytic polysaccharide monooxygenase mutant provided by the invention has an application prospect in the aspects of degrading biomass or plant polysaccharide and the like.

The invention relates to the technical field of molecular biology, in particular to a qPCR (quantitative polymerase chain reaction) detection kit for detecting cis-trans mutation and a use method of the qPCR detection kit. The invention provides a primer pair which is used for detecting a first mutation site and / or a second mutation site of a target gene and comprises a universal primer F matched with the upstream of the first mutation site, a downstream primer R-BW matched with a wild type sequence of the second mutation site and a downstream primer R-BM matched with a mutation type sequence of the second mutation site, the fluorescent probe P-AW is matched with the upstream of the first mutation site, the fluorescent probe P-AM is matched with the mutant sequence of the first mutation site, and the fluorescent groups of the fluorescent probe P-AW and the fluorescent probe P-AM are different. According to the reaction system disclosed by the invention, the MGB fluorescent probe and the ARMS primer are combined, and mutation information and a cis-trans relationship can be judged according to the existence of a fluorescent signal. Compared with an existing detection method, the method for detecting the cis-trans relation through fluorescent quantitative PCR is proposed for the first time, detection can be completed within 2 h, specificity is good, the cost advantage is achieved, operation is easy and convenient, and clinical operation is convenient.

The invention discloses a phytolacca americana seed EMS mutation library establishment method, and belongs to the technical field of mutation library construction.The phytolacca americana seed EMS mutation library establishment method comprises the steps that concentrated sulfuric acid is used for pretreating phytolacca americana seeds, then germination accelerating is conducted on the seeds, and the seeds are treated with an EMS solution when the seeds are white, so that the problems that phytolacca americana seed coats are thick, and the sensitivity to EMS is low are solved; according to the mutagenesis method disclosed by the invention, high-density series allele point mutation can be induced and generated, the mutagenesis frequency is relatively high, and meanwhile, chromosome distortion is less caused. In the mutagenesis process, abundant genetic variation can be more efficiently created, and more possibilities are provided for screening mutants with excellent characters. Besides, the method has a remarkable advantage in the aspect of cost, the required chemical reagent is cheap, the experimental operation process is simple, and complicated equipment and technology are not required like common irradiation. The breeding cost is reduced, the working efficiency is improved, and wide popularization and application are facilitated.

The invention discloses a recombinant protein for detecting an anti-myelin sheath related glycoprotein MAG antibody and a detection reagent of the recombinant protein, and belongs to the technical field of neural immunodiagnostics, five amino acid sites in MAG protein are subjected to specific amino acid point mutation replacement to obtain an MAG protein mutation sequence, the MAG protein mutation sequence is fused with GlcATP enzyme and HNK-1ST enzyme full-length sequences to construct a lentiviral expression vector, and the recombinant protein and the detection reagent are used for detecting the anti-myelin sheath related glycoprotein MAG antibody. According to the present invention, three genes are constructed, and are stably transfected in a host cell so as to stably express the three genes in one cell at the same time, such that the constructed recombinant protein and the wild type MAG protein have similar anti-MAG IgM antibody binding domain, and on the premise of not using GlcATP enzyme and HNK-1ST enzyme small molecule agonists, the sensitivity and the specificity of the detection of the MAG autoimmune antibody by using the CBA method are improved; the method is suitable for preparing related detection reagents for in-vitro detection of the IgM type anti-MAG antibody.