32 results about "Conserved sequence" patented technology

The invention provides a primer for detecting a novel bunyavirus by an RPA-lateral flow nucleic acid chromatography technology, which comprises a specific SFTSV primer pair marked by a nucleic acid sequence, namely SFTSV-F and SFTSV-R. The invention further provides a kit for detecting the novel bunyavirus by the RPA-lateral flow nucleic acid chromatography technology. The invention also provides a probe which comprises a gold nanoparticle capture probe, an SFTSV capture probe and a quality control capture probe. Meanwhile, the invention further provides application of the primer and the probe in preparation of SFTSV detection products, a specific detection kit and a specific detection method. The specific primer and the functional differentiation probe are designed aiming at a novel bunyavirus highly-conserved sequence, and an RPA isothermal amplification technology and a lateral flow nucleic acid chromatography visual detection technology are integrated, so that the rapid, sensitive and specific detection of the SFTSV is realized, the dependence on a large instrument is not needed, the detection cost and the operation threshold are reduced, and the detection efficiency is improved. And the application requirements of on-site rapid detection, large-scale screening and resource-limited regions are met.

This invention provides a method for identifying abamectin resistance genes in the fall armyworm, dsRNA, and their applications, belonging to the field of molecular biology. This invention combines genome resequencing and transcriptome sequencing to identify key abamectin resistance genes in the fall armyworm. Based on the conserved mRNA sequences of the key resistance genes GSTD6 and CTP6A18, dsRNA was designed, and RNAi technology can effectively reduce the fall armyworm's resistance to abamectin. This has significant practical implications for developing effective fall armyworm resistance management strategies to delay the emergence of resistance and for identifying new targets for pest control.

This invention belongs to the field of bioengineering technology and discloses a method for specific targeting and scarless simultaneous elimination blue KPC‑2 Gene and IncL plasmid sgRNA sequences, CRISPR / Cas9 vector system and applications. This system targets the sgRNA of the IncL plasmid. blue KPC‑2 The IncL-type plasmid of the gene was designed with an sgRNA that specifically recognizes the conserved sequence at its origin of replication, thus eliminating the drug resistance gene while disrupting the plasmid's replication ability. Conjugation transfer to recipient bacteria significantly improved vector delivery efficiency. An arabinose-inducible promoter was used to control Cas9 gene expression, improving system stability and editing success rate. The vector carries a sucrose-inducible self-eliminating gene, enabling controlled vector self-elimination. This system is not a simple superposition but rather constitutes a time-controlled operational flow: efficient delivery via conjugation, controlled cleavage via induction, and finally, induced self-elimination.

The invention relates to the technical field of construction of conditional gene knockout models, and discloses a construction method of a mouse Oct4 conditional gene knockout model based on quadruple sgRNA collaborative targeting, which comprises the following steps: step 1, designing four sgRNAs, namely sgRNA1, sgRNA2, sgRNA3 and sgRNA4, in which the sgRNA1 and the sgRNA2 target 5'end and 3 'end non-coding regions of a second exon of an Oct4 gene, and the sgRNA3 and the sgRNA4 target structural domain conserved sequences; step 2, preparing an electrotransfection solution, wherein the electrotransfection solution is composed of Cas9mRNA, the four sgRNAs and an electrotransfection buffer solution; step 3, carrying out electroporation treatment on the fertilized eggs of the mice; step 4, culturing the fertilized eggs subjected to the electrotransformation treatment to a blastocyst stage; and 5, carrying out genotype identification on the cultured blastocyst. According to the scheme, the sgRNA combination and embryo electrotransfection system is optimized, the mouse Oct4 conditional gene knockout model is efficiently and accurately constructed, a reliable tool is provided for regulation mechanism research of Oct4 in mouse early embryo trigerm differentiation, and the medical transformation value of Oct4 is excavated.

The invention provides a fluorescent quantitative PCR (polymerase chain reaction) detection primer, a kit and a method for a novel variable chalmavirus VP gene, and belongs to the technical field of virus detection. A specific primer is designed and synthesized according to a conserved sequence of a VP gene of the novel variant chalmavirus, a recombinant plasmid is constructed, then the recombinant plasmid is used as a standard substance to establish a standard curve, a real-time fluorescent quantitative PCR detection system aiming at the novel variant chalmavirus is constructed, and specific detection of the novel variant chalmavirus is realized. The detection method can be used for quantitatively detecting the novel variant chalmavirus in the to-be-detected sample, and has the characteristics of high speed, high sensitivity, strong specificity and the like; the method provides a key technical support for molecular epidemiological investigation of the Sepamavirus in animal populations, is helpful for realizing early screening of infected individuals and timely blocking of virus transmission, and has important practical significance for guaranteeing population health and maintaining high mountain ecosystem stability.

The invention provides a real-time fluorescent quantitative PCR (polymerase chain reaction) detection primer, kit and method for novel variant bocavirus, and belongs to the technical field of virus nucleic acid detection. A specific primer is designed and synthesized according to a conserved sequence of a novel variant bocavirus (BoV) VP gene, a recombinant plasmid pcDNA3.1 HA-BoV-VP is constructed, then the recombinant plasmid is used as a standard substance to establish a standard curve, a real-time fluorescent quantitative PCR detection system for the novel variant bocavirus is constructed, and specific detection of the novel variant bocavirus is realized. The detection method can realize quantitative detection of bocavirus in a to-be-detected sample, has the characteristics of high speed, high sensitivity, strong specificity and the like, plays an important role in protection of endangered wild animals such as disc sheep, and has good practicability.

The invention discloses a primer combination for detecting Ralstonia pickettii and a kit for detecting the Ralstonia pickettii, wherein the kit comprises the primer combination. The primer probe provided by the invention is targeted to a specific nucleotide sequence of the Ralstonia piceatrii instead of a horizontal conserved sequence, so that the Ralstonia piceatrii and other strains in the Ralstonia can be effectively distinguished, the technical defects that an existing detection method is insufficient in specificity and easy to generate false positive are overcome, and the accuracy of a detection result is improved. The detection limit can reach 100 CFU / mL, and compared with the prior art, the detection limit is reduced, and the detection sensitivity is improved. The kit disclosed by the invention can be applied to pollution detection of medical supplies such as pharmaceutical water, dialysis water, endoscope cleaning water, blood products, sterile normal saline and the like and rapid screening of hospital infection, is strong in specificity and high in accuracy, and has relatively high popularization and application values.

The invention provides a ZmSWEET4 gene related to a corn smut disease and application of the ZmSWEET4 gene in regulation and control of the corn smut disease, and belongs to the technical field of gene engineering. The invention provides a ZmSWEET4 gene capable of regulating and controlling the resistance of corn to the tumor smut disease. A coded protein contains conserved sequences as shown in SEQ ID No.1 and SEQ ID No.2. The invention also provides a preparation method of the ZmSWEET4 gene capable of regulating and controlling the resistance of corn to the tumor smut disease. According to the present invention, the ZmSWEET4a gene and the ZmSWEET4b gene in the receptor corn are respectively edited to obtain the transgenic plant in which the ZmSWEET4a gene and the ZmSWEET4b gene are respectively knocked out, and compared with the wild type corn, the gene knockout plant has characteristics of more disease resistance. The invention proves that the ZmSWEET4 gene is related to the resistance of corn to Ustilago esculenta for the first time, and can be used for creating new germplasm resistant to the corn Ustilago esculenta and developing or screening medicines capable of preventing and / or treating the corn Ustilago esculenta.

The invention belongs to the technical field of biological genetic engineering, and discloses a CRISPR / Cas9 (clustered regularly interspaced short palindromic repeats / associated protein 9) system for specific targeted traceless elimination of a blaNDM-1 gene and an IncN plasmid and application of the CRISPR / Cas9 system. According to the system, sgRNA of an original point conserved sequence is copied by designing an IncN type plasmid carrying a blaNDM-1 gene in a targeting manner, so that the replication capability of the plasmid is destroyed while a drug-resistant gene is eliminated; the carrier delivery efficiency is greatly improved by conjugational transfer to recipient bacteria; the expression of the Cas9 gene is controlled by adopting an arabinose inducible promoter, so that the constitutive toxicity of Cas9 protein is reduced, and the system stability and the editing success rate are improved; the sucrose-induced carrier self-elimination gene carried by the carrier realizes controllable self-elimination of the carrier. The system is not simply superposed, but forms an operation process with a controllable time sequence: efficient delivery is firstly carried out through joint, then cutting is controlled through induction, and finally self-elimination is induced.

The present application relates to the technical field of plant gene function, and aims to provide a construction method of a homologous silencing system of a Paeonia lactiflora bud dormancy gene, comprising the following steps: designing amplification primers according to gene annotation and a conserved sequence region in a Paeonia lactiflora bud dormancy full-length transcriptome, and constructing a conserved sequence fragment of a target gene into Agrobacterium tumefaciens containing a pTRV vector; taking a rhizome of a treated Paeonia lactiflora annual seedling as an infection material, and placing the infection material in Agrobacterium tumefaciens liquid containing pTRV1, pTRV2 and pTRV2-target gene fragments to perform infection treatment; potting the rhizome and moving the rhizome into a culture room to perform culture, observing phenotype and expression amount differences between the rhizome and a control group in a growth process, and confirming that the homologous silencing system is successfully constructed. The present application first completes miniaturization treatment and batch bacterial liquid infection of an underground rhizome system of Paeonia lactiflora, establishes a virus-mediated homologous function silencing system of a Paeonia lactiflora bud dormancy gene, and realizes the purpose of verifying functions of bud dormancy and even germination and growth related genes.

The invention discloses a recombinant protein as well as a coding gene and application thereof, and belongs to the technical field of bioengineering. The invention designs a recombinant protein which is composed of two subunits with different functions, and design and transformation are carried out on the basis of a conserved sequence of an alpha herpes virus protein with an immune escape function, so that the secretory expression of the recombinant protein is promoted, and the defects of inclusion body formation, overhigh host toxicity and high target protein purification difficulty are overcome. Experimental results show that the recombinant protein provided by the invention has a remarkable effect of inhibiting innate immunity of cells, and can effectively inhibit interferon generated by innate immunity of cells, so that the titer of cultured viruses is improved.

The application belongs to the field of biological detection, and particularly relates to an isothermal amplification kit for detecting porcine circovirus type 3. The application provides an isothermal amplification detection primer group for porcine circovirus type 3 (PCV3), which comprises a pair of outer primers (LOF and LOR) and a pair of inner primers (LIF and LIR) and is designed according to the conserved sequence of the porcine circovirus type 3. The primer group has low design difficulty, and maintains similar sensitivity and specificity to other isothermal amplification reactions such as a loop-mediated isothermal amplification method, can be suitable for target sequences with higher GC content and below 200 bp, has no denaturation and annealing process, is suitable for on-site detection, and has great application potential.

The invention provides a detection method for detecting Sf9 host DNA residues by using a fluorescent probe and application, and belongs to the technical field of biological detection.According to the method, a specific primer pair (such as SEQIDNO: 1 / 2) and the fluorescent probe (such as SEQIDNO: 11) are designed according to a high-copy conserved sequence of Sf9 host DNA, a kit containing a gene amplification solution, a standard substance and a quality control substance is matched, DNA is extracted through a liquid nitrogen grinding-column extraction method, and the detection result is accurate. And carrying out real-time fluorescent quantitative PCR amplification and then carrying out detection. According to the method, the detection sensitivity reaches 0.003 pg / mu L, the linear range is 300 pg / mu L-0. 003 pg / mu L, cross reaction to irrelevant DNA is avoided, the repeatability CV% is smaller than or equal to 10%, the reagent is still stable after being frozen and thawed for 10 times, and the whole process is completed within 1.5 hours; the method solves the problems of insufficient specificity and sensitivity, tedious operation and the like of the existing method, and can be used for quality control of biological products such as gene therapy drugs and cell vaccines.

The invention discloses a multiple qPCR (quantitative polymerase chain reaction) detection composition, method and kit for detecting various DNA / RNA (deoxyribonucleic acid / ribonucleic acid) viruses, and aims to detect various human DNA / RNA viruses possibly existing in biological products, compare and analyze through database analysis, find virus genome conserved sequences, design primers and probes with higher specificity and detect most subtypes of the viruses. A plurality of pairs of probe primers are designed for a virus conserved region, the primer probes with good specificity and high sensitivity are finally screened, the virus content of 2 * 10 < 7 > copies / [mu] L to 20 copies / [mu] L can be accurately quantified, and the sensitivity of most viruses can reach 2-10 copies / [mu] L. By repeatedly screening and optimizing a reaction system, including the concentrations of the primers and the probes, the detection method can be used for stably detecting various DNA / RNA viruses under the condition of fewer nucleic acid samples, is simple to operate and has higher sensitivity.

The application discloses a triplex forming oligonucleotide fluorescent probe and a detection method for detecting mycoplasma by targeting P1 adhesion protein genes, and belongs to the technical field of microbial molecular detection. The fluorescent probe is a triplex forming oligonucleotide (TFO), targets a specific conservative sequence of mycoplasma P1 adhesion protein genes, the 5' end of the fluorescent probe is modified with a fluorescent reporter group, the 3' end of the fluorescent probe is modified with a fluorescent quenching group, the fluorescent probe can form a stable triplex DNA complex with the target sequence through Hoogsteen base pairing and generate a fluorescent signal. The detection method comprises four core steps of sample pretreatment and DNA extraction, fluorescent probe incubation, fluorescent signal detection and result judgment, and does not need PCR amplification, and is simple in operation and rapid in detection. The probe has high specificity and high sensitivity, the detection lower limit can reach 1 copy / muL, is suitable for mycoplasma detection of various types of samples, and can be widely applied to scenes such as clinical diagnosis, biological pharmaceutical quality control and livestock breeding quarantine.

This invention discloses a PSR primer, kit, and detection method for identifying Pediococcus lactis based on a specific gene target. Based on the conserved sequence of the target gene, this invention designs and optimizes PSR-specific primers, establishes a PSR amplification system, and further provides a kit containing the primers and its application method. The PSR detection method established by this invention features high sensitivity, strong specificity, rapid reaction, simple operation, and low cost. It is suitable for the rapid and accurate identification of Pediococcus lactis and has promising application prospects in food fermentation, probiotic detection, and microbial quality control.

The invention discloses a primer group for identifying and distinguishing mycobacterium tuberculosis and nontuberculous mycobacterium, the primer group is a pair of RPA (recombinase polymerase amplification) primers for specifically amplifying a target gene of the mycobacterium tuberculosis, and the target gene is a conserved sequence rpoB gene of the mycobacterium tuberculosis. Accordingly, the inventor also develops a corresponding composition and a kit. On the basis, a method for identifying and distinguishing mycobacterium tuberculosis and nontuberculous mycobacterium based on RPA-CRISPERCas12a for a non-diagnostic purpose is established, firstly, a sample to be detected is subjected to RPA amplification, and the primer group is adopted for RPA amplification; and then, carrying out CRISPR / Cas12a detection on the RPA amplification product by adopting the crRNA and the CRISPR / Cas12a detection reagent. The method has the advantages of low cost, high practicability and simplicity and convenience in operation, is suitable for quickly screening or identifying and distinguishing the mycobacterium tuberculosis and the four non-tuberculosis mycobacteria on site, and has a wide application prospect.

The invention discloses an Eucommia virus induced gene silencing system as well as a construction method and application thereof. The system is obtained by infecting Eucommia ulmoides with a target gene fragment recombinant virus plasmid carrying a target gene through agrobacterium tumefaciens; a target gene is an Eucommia ulmoides phytoene dehydrogenase gene, namely an Eucommia ulmoides EuPDS gene, and the nucleotide sequence of the EuPDS gene is as shown in SEQ ID NO. 1; the target gene segment is a conservative CDS sequence on the EuPDS gene of Eucommia ulmoides, and the nucleotide sequence of the target gene segment is as shown in SEQ ID NO.2. The Eucommia virus-induced gene silencing system can be used for studying Eucommia EuPDS gene functions, Eucommia phytoene dehydrogenase metabolic pathways or Eucommia stress resistance, and has profound significance for studying the Eucommia gene functions.

The invention discloses a qPCR (quantitative polymerase chain reaction) method for quantitatively analyzing human cells in a non-humanized animal model, which comprises the following steps: taking a non-humanized animal model sample, and extracting genome DNA (deoxyribonucleic acid); the method comprises the following steps: designing and synthesizing a specific primer probe combination aiming at a human COX1 gene conserved sequence; the method comprises the following steps: cloning a specific fragment of a human COX1 gene to a plasmid vector, and preparing a linearized plasmid standard substance; carrying out 10-fold gradient dilution on the linearized plasmid standard substance to obtain a standard substance gradient solution, and fitting a linear regression curve; taking the extracted DNA as a template, adding a primer probe combination and a qPCR premixed solution, and carrying out qPCR amplification; and calculating the human cell content in the sample according to the linear regression curve. The method provided by the invention has the advantages of stable standard substance, strong species specificity, high detection sensitivity, cross-species universality and simple operation, can effectively avoid animal matrix DNA interference, solves the problems of quantitative deviation and low-abundance cell detection, and reduces the detection cost.

The application is designed and screened according to the specific amplification primer of the conserved sequence of Chinese mitten crab microsporidium, the specific amplification primer screened is modified with a marker, and then the MIRA reaction condition is optimized; the MIRA detection technology has obvious advantages such as no need for thermal cycling, short detection time, simple operation, high sensitivity, simple equipment configuration and the like; compared with ordinary PCR detection, the MIRA detection technology of Chinese mitten crab microsporidium established in the experiment can complete the reaction at 37 DEG C in only 25 min, the minimum detection limit is 1 copies / µL, which is 10 4 copies / µL higher than that of ordinary PCR, the detection accuracy reaches 100%, and good specificity is achieved.

The invention provides a non-invasive PCR (Polymerase Chain Reaction) sex identification method for tortoises, and belongs to the technical field of tortoise sex identification. The method comprises the following steps: extracting genome DNA from a non-invasive sample, taking the genome DNA as a template, carrying out PCR amplification by using primers as shown in SEQ ID NO.1-6, and judging that the non-invasive sample is male when an internal reference band of 432 bp and a male specific band of 278 bp appear in an amplification product at the same time; and when an internal reference band of 432 bp and a female specific band of 196 bp simultaneously appear in the amplification product, judging that the amplification product is female. Amplification primers used in the method are designed according to conserved sequences of gender-specific genes of the tortoises, the primers are extremely high in specificity, do not have cross amplification with reptiles such as snakes and lizards, and are adaptive to more than 10 common tortoises such as grass tortoises, brazilian tortoises and land tortoises, the specificity reaches 100%, the accuracy reaches 99% or above, and the method has good popularization and application prospects.

The invention discloses an accurate sequence analysis method and system based on nanopore length and read length sequencing data. The method comprises the following steps: comparing nanopore sequencing data with a reference genome through a minimap2 algorithm, and screening out high-quality long-read-length sequences in a target area and specific upstream and downstream ranges; extracting a target area local sequence of each read length, calculating an editing distance matrix, performing Box-Cox transformation, performing clustering analysis by adopting a Gaussian mixture model, and determining an optimal clustering number through a BIC criterion; performing multi-sequence alignment and polishing on each type of sequences to generate a high-precision consensus sequence; and finally, recognizing homologous conserved sequence fragments through re-comparison and global comparison, and outputting candidate sequences for targeting PCR (Polymerase Chain Reaction) primer or CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) sgRNA design. According to the method, the problem of high error rate of nanopore length and read length data can be effectively solved, the accuracy and reliability of sequence analysis are improved, and reliable technical support is provided for precise gene editing and molecular diagnosis.

A bacteria test method according to the invention tests for the presence or absence of bacteria in a sample, and includes amplifying a conserved sequence in bacterial 23S rRNA by PCR using a primer set consisting of a primer (1) and a primer (2). Primer (1): A primer having a sequence of SEQ ID NO: 2, and / or a primer having a sequence in which one to three bases have been modified in the sequence of SEQ ID NO: 2. Primer (2): At least one selected from the group consisting of primers having a sequence of SEQ ID NO: 3, primers having the sequence of SEQ ID NO: 3 in which one to three bases have been modified, primers having a sequence of SEQ ID NO: 36, primers having the sequence of SEQ ID NO: 36 in which one to three bases have been modified, primers having a sequence of SEQ ID NO: 39, primers having the sequence of SEQ ID NO: 39 in which one to three bases have been modified, primers having a sequence of SEQ ID NO: 41, and primers having the sequence of SEQ ID NO: 41 in which one to three bases have been modified.

The present invention provides compositions and methods for making circular RNAs, in particular circular RNAs encoding one or more proteins and having superior stability. Such circular RNAs comprise complementary regions that are capable of forming double-stranded regions that are important for maintaining the stability of the circular RNA under different environments and that facilitate translation and circularization. Such complementary regions can be formed by codon optimization or can be introduced as heterologous sequences. The present invention also provides circular RNAs that express self-amplifying RNA non-structural proteins, e.g., (nsP1-4), which can amplify independent mRNA molecules containing corresponding conserved sequence elements (CSEs). These mRNAs can encode a gene of interest (GOI). When they are introduced together into a target cell, the circular RNAs can amplify the mRNAs, thereby enabling stable high expression of the gene of interest.

The invention discloses an avian leukosis virus subgroup rapid detection kit and a detection method thereof, and belongs to the technical field of animal inspection and quarantine. Aiming at the defects of the existing detection technology, a conserved sequence of an env gene shared by A, B and K subgroups and a conserved sequence of a specific env gene of a J subgroup are selected as target regions, specific primer probes are designed, a reaction system and an amplification program are optimized, and the kit is assembled. The kit adopts a fluorescent RT-PCR (Reverse Transcription-Polymerase Chain Reaction) method, can simultaneously detect A, B, J and K subgroups and accurately distinguish endogenous and exogenous viruses without cross interference. The kit has the characteristics of high detection speed, high sensitivity, strong specificity, simplicity and convenience in operation and the like, is suitable for detecting various samples such as chicken plasma, seminal fluid and cloaca swab, can improve the breeding chicken purification efficiency, shorten the period and reduce the economic loss of the poultry industry, and has remarkable application value.

The application provides a H1N1 broad-spectrum mRNA vaccine based on key amino acid mutation of HA and NA protein series as a target and a preparation method thereof, and relates to the technical field of vaccine preparation.The amino acid sequence of the mRNA vaccine is shown as SEQ ID NO.1, and the nucleic acid sequence is shown as SEQ ID NO.2, that is, on the basis of the HA conservative sequence, variation sites of amino acid properties across the vicinity of the main antigen site of the HA head and the receptor binding site are retained, the HA' amino acid sequence containing the key mutation site is obtained, and the HA' and the NA conservative amino acid sequence are connected by GGGSGGGSGGGSGGGS to obtain the mRNA vaccine.The application overcomes the defects of the prior art, improves the broad-spectrum protection effect on the H1N1 type influenza virus by retaining the key amino acid mutation site in the HA conservative sequence, reduces the vaccine production cost and the immunization cycle, and provides an effective response scheme for the influenza outbreak season.

The application discloses a method and device for identifying CNEs (Conserved Non-coding Elements) in a genome. The method comprises: aligning each of a plurality of high-quality genomes with a reference genome to obtain genome alignment results; identifying a species evolution relationship topological structure and the genome alignment results to obtain a conserved sequence position information gff file; filtering the gff file to obtain a CNEs.gff file; extracting CNEs sequences from the CNEs.gff file; determining CNEs differentiation sequences according to the CNEs sequences; and performing functional enrichment analysis on target genes of the CNEs differentiation sequences to obtain CNEs identification results in the plurality of high-quality genomes. The application solves the technical problem in the related art that the identification of CNEs is often affected by genome structural variations, resulting in incomplete identification of CNEs or misjudgment.

The invention belongs to the field of agricultural microorganism application, and particularly relates to a primer group, a reagent or a kit, a strain screening method adopting the primer group, the reagent or the kit and application, and the primer group is designed according to a conserved sequence of a structural gene segment in a gene cluster synthesized by iturin A; wherein the structural genes in the iturin A synthetic gene cluster comprise an ituA gene, an ituB gene and an ituC gene; the primer group comprises a primer pair I, a primer pair II and a primer pair III. The primer group disclosed by the invention is designed according to a conserved sequence of a structural gene segment in an iturin A synthetic gene cluster, so that key structural gene segments (ituA gene, ituB gene and ituC gene) in the iturin A synthetic gene cluster can be specifically amplified, and by using the primer group and combining a PCR (Polymerase Chain Reaction) amplification technology and an electrophoresis detection technology, the detection sensitivity of the iturin A synthetic gene cluster is improved, and the detection sensitivity of the iturin A synthetic gene cluster is improved. The bacillus positive strain capable of producing the iturin A can be rapidly and accurately screened at low cost, and the application prospect is wide.

The invention discloses recombinant corynebacterium glutamicum capable of efficiently expressing heterologous protein and application of the recombinant corynebacterium glutamicum, and belongs to the field of genetic engineering and bioengineering. The invention constructs a T7 expression system taking corynebacterium glutamicum as a chassis cell, and applies the T7 expression system to expression of various heterologous protein genes. The T7 expression system designed by the invention is not inhibited by glucose metabolism, and the expression effect of the T7 expression system in a culture medium with glucose is obviously superior to that of a common system. The invention also finds that a combination of a T7 promoter and a section of special RBS sequence has an expression effect obviously better than that of a common conserved RBS sequence of corynebacterium glutamicum.

A fungal test method according to the present invention is for testing whether a fungus is present in a sample, and includes amplifying a conserved sequence within fungal 25S / 28S rRNA by PCR using a primer set comprising a primer (1) and a primer (2). Primer (1): a primer having a sequence of 13-25 bases including the sequence of SEQ ID NO: 2, and / or a primer having a sequence of 10-25 bases including a sequence obtained by modifying 1-3 bases in the sequence of SEQ ID NO: 2. Primer (2): a primer having the sequence of SEQ ID NO: 3, and / or a primer having a sequence obtained by modifying 1-3 bases in the sequence of SEQ ID NO: 3.