31 results about "Gene clone" patented technology

This invention discloses a SUMO-C7H 15 This invention relates to the field of biotechnology, specifically to NO2S probes, their synthesis methods, and applications. The molecular formula of the probe described in this invention is SUMO-C7H. 15 NO2S, wherein the SUMO is human SUMO-1; the nucleotide sequence of the human SUMO-1 is shown in SEQ ID NO: 1, and the amino acid sequence is shown in SEQ ID NO: 2. This invention obtains the polypeptide sequence encoding the human SUMO-1 protein through gene cloning technology, and further prepares a structurally stable SUMO-C7H protein using chemical synthesis methods. 15 NO2S, this probe has a stable molecular structure and is easy to synthesize and detect; SUMO-C7H 15 The NO2S fluorescent probe possesses high specificity and good detection sensitivity, providing an innovative research tool for exploring tumorigenesis mechanisms and assessing SUMOylation modification levels.

This invention relates to the field of bioengineering technology, and more particularly to a gene for identifying salt tolerance in upland cotton, GhNRX16, and its molecular marker. The CDS sequence of the upland cotton gene GhNRX16 is shown in SEQ ID NO.1 of the sequence listing. This invention uses RNA-seq technology to screen and identify this salt tolerance-related gene in the transcriptome data of upland cotton under salt stress treatment. Furthermore, the GhNRX16 sequence was obtained through gene cloning, and a transient silencing vector based on VIGS (virus-induced gene silencing) technology was successfully constructed. Experiments using VIGS-mediated gene silencing combined with salt stress treatment showed that silencing the GhNRX16 gene significantly reduced the plant's tolerance to salt stress, demonstrating that normal expression of this gene directly contributes to ensuring the plant's salt stress resistance. These results confirm the crucial role of the GhNRX16 gene in regulating cotton salt tolerance, providing important gene resources and theoretical basis for molecular breeding of salt-tolerant cotton.

This invention relates to the field of crop genetics and breeding, and discloses the sweet sorghum disease resistance gene CPK4, its encoded protein, and its applications. The nucleotide sequence of the sweet sorghum disease resistance gene CPK4 is shown in SEQ ID NO.1, or has more than 90% homology with the sequence shown in SEQ ID NO.1 and encodes a protein with a nucleotide sequence that promotes the release of reactive oxygen species (ROS) in plants. By utilizing the CPK4 gene cloned from sweet sorghum, and through the specific interaction between its encoded protein and the NADPH oxidase SbRbohD protein, the ROS signaling pathway induced by pathogens is specifically activated, strengthening the defense response of sweet sorghum against anthracnose. This achieves a targeted enhancement of sweet sorghum's resistance to anthracnose. Furthermore, this gene is specifically induced by the molecular patterns of anthracnose and pathogens, and its disease resistance mechanism differs from existing NLR-mediated immune pathways, enriching the types of sweet sorghum disease resistance gene resources.

This invention discloses a Panax notoginseng ethylene response factor gene. PnERF120 Its nucleotide sequence is shown in SEQ ID NO:1, and the encoded amino acid sequence is shown in SEQ ID NO:2. PnERF120 The gene originates from Panax notoginseng, and the encoded protein sequence contains a typical conserved domain of the ERF transcription factor. This invention demonstrates, through gene cloning, bioinformatics analysis, expression pattern analysis, subcellular localization, overexpression, and other techniques and methods, that... PnERF120 This invention can be used to improve the resistance of Panax notoginseng to Panax notoginseng A virus, and also to improve the resistance of tobacco to tobacco mosaic virus. This invention provides new functional genes and application pathways for antiviral breeding and disease-resistant materials of Panax notoginseng and other plants.

The present application relates to the technical field of biological genetic engineering, and particularly relates to a kind of identification of plant salt tolerance land cotton gene GhNRX71 And molecular marker, the CDS sequence of the land cotton gene GhNRX71 As shown in SEQ ID NO.1 In the sequence table.The present application is screened and identified in the transcriptome data of land cotton under salt stress treatment by transcriptome sequencing (RNA-seq) technology.The sequence of GhNRX71 Is obtained by further gene cloning means, and the transient silencing vector based on VIGS (virus-induced gene silencing) technology is successfully constructed.The results show that the tolerance of plant to salt stress significantly reduces after silencing GhNRX71 Gene by using VIGS-mediated gene silencing combined with salt stress treatment experiment, which proves that the normal expression of the gene has a direct effect on guaranteeing the salt stress resistance of plant.The above results confirm that GhNRX71 Gene plays a key role in regulating the salt tolerance of cotton, and provides important gene resources and theoretical basis for cotton salt-tolerant molecular breeding.

The application discloses a CMV virus-like particle for producing a VLP recombinant vaccine and a preparation method thereof, the CMV virus-like particle is obtained by cloning a C-terminal G4SLPETG modified plant virus cucumber mosaic virus CMV capsid gene into a prokaryotic expression vector to obtain a recombinant expression vector, the recombinant expression vector is transfected into E. coli BL21 (DE3), and expression is obtained through the recombinant E. coli BL21 (DE3); and the amino acid sequence of the C-terminal G4SLPETG modified CMV is SEQ ID NO. 1. Test proves that the recombinant strain constructed by the application is stable to expression of an exogenous protein. The recombinant protein expressed by the application is used for preparing a virus-like particle for antigen coupling, the coupling efficiency is high, the antigen purity is high, the safety is good, the virus-like particle is not pathogenic to mice and other animals, and the safety evaluation is easy.

This invention discloses a ribonuclease derived from tartary buckwheat. FtRNH Genes, their expression vectors, and applications. This invention isolates ribonuclease from tartary buckwheat through gene cloning. FtRNH The gene, whose CDS sequence is shown in SEQ ID No. 1, and whose encoded protein amino acid sequence is shown in SEQ ID No. 2, is described in this invention. FtRNH Transgenic lines were obtained by genetic transformation into cultivated tartary buckwheat (ZK3) and subjected to ultraviolet stress treatment. Compared with wild-type materials, the transgenic lines showed increased expression. FtRNH The positive lines of tartary buckwheat exhibited stronger tolerance to ultraviolet radiation and less accumulation of UV-B-induced R-loops and cyclobutanepyrimidine dimers. This invention has significant application prospects in improving the tolerance of cultivated tartary buckwheat to ultraviolet stress or in breeding tartary buckwheat varieties resistant to ultraviolet stress.

This invention belongs to the fields of genetic engineering and enzyme engineering, specifically relating to a chitinase mutant ChiTgM and its applications. This invention obtains the chitinase ChiTg through gene cloning, achieves heterologous expression of it in Pichia pastoris through codon optimization, and obtains the chitinase mutant ChiTgM through site-directed mutagenesis and combinatorial mutagenesis. Analysis of the enzymatic characteristics of chitinase ChiTg and the mutant ChiTgM shows that, compared to the starting template ChiTg, the chitinase mutant ChiTgM provided by this invention exhibits a 1.81-fold increase in specific activity and demonstrates excellent hydrolytic properties. Its application in the enzymatic preparation of chitin oligosaccharides can efficiently hydrolyze colloidal chitin, laying the foundation for its further industrial application.

The invention belongs to the technical field of genetic engineering and biology, and particularly relates to a method for rapidly extracting high-quality genome DNA from thick-wall microalgae and application. The invention optimizes a set of rapid genome DNA extraction method by splitting chlorella cells through helicase combined with vacuum treatment. Compared with a cellulase combined vacuum treatment method and two commercial genome DNA extraction kits, the genome DNA obtained by the method is higher in integrity. Besides, the genome DNA extracted by the optimization method is completely suitable for downstream applications such as gene cloning and genotyping, the extraction integrity of the genome DNA of chlorella is remarkably improved, and a new thought can be provided for high-quality genome DNA extraction research of other types of microalgae.

This invention discloses an anti-ERBB2 monoclonal antibody, its preparation method, and its applications, belonging to the field of biomedical technology. This anti-ERBB2 monoclonal antibody uses the intracellular domain of the human ERBB2 protein as an immunogen and is obtained through animal immunization, B cell screening, gene cloning, and recombinant expression techniques. It can specifically recognize the human ERBB2 protein, exhibiting high affinity and high specificity, and can recognize both native and denatured ERBB2 proteins. This monoclonal antibody can be applied in immunohistochemistry, immunofluorescence, Western blotting (WB), flow cytometry, ELISA detection, and tumor targeted therapy. Experimental results show that this antibody has low background and no non-specific binding, accurately detecting ERBB2-overexpressing tumor cells and tissues, and has significant application value in the diagnosis, prognostic assessment, and targeted therapy of ERBB2-related diseases such as breast cancer.

The application discloses a DREB transcription factor CtDREB1B of safflower and application thereof in positive regulation of drought and salt stress resistance of plants. The full-length nucleotide sequence of the gene CtDREB1B is shown as SEQ ID NO. 2, and the amino acid sequence of the encoded protein is shown as SEQ ID NO. 1. The application first clones the complete coding frame of the gene by RT-PCR, clones the gene into a pYES2 vector and transfers the gene into yeast, and finds that the pYES2::CtDREB1B yeast cells can still grow on SGR culture medium containing 1.5 M mannitol or 4 M NaCl. Then, the CtDREB1B gene is heterologously expressed into Arabidopsis thaliana. Overexpression of the CtDREB1B in the Arabidopsis thaliana can make the transgenic Arabidopsis thaliana exhibit a drought and salt tolerance phenotype, and the activities of protective enzymes such as SOD and POD are enhanced, and the expression amounts of stress resistance related genes such as AtABF3 and AtRD29A are up-regulated, indicating that the gene can simultaneously positively regulate drought and salt stress. Therefore, the gene can be applied to breeding of new plant stress resistance varieties, and in addition, the gene provides a new gene resource for plant stress resistance, and has important application prospects in breeding of salt-tolerant plants.

This invention discloses the chitin synthase 1 (LmCHS1) gene of the leafminer moth, its dsRNA, its synthesis method, and its insect-resistant applications. Through transcriptome analysis, specific primer pairs for the open reading frame of the LmCHS1 gene were designed and synthesized. The LmCHS1 gene sequence was obtained through gene cloning (nucleotide sequence: SEQ ID NO: 1, amino acid sequence: SEQ ID NO: 2). The gene fragment with sequence SEQ ID NO: 3 was selected as the RNAi target sequence. Primer pairs SEQ ID NO: 6 and SEQ ID NO: 7 with restriction enzyme sites were designed and synthesized. PCR amplification was performed, and a prokaryotic expression vector was constructed to induce dsRNA expression. Feeding leafminer moth larvae with dsRNA interfered with LmCHS1 gene expression. After 24 hours, abnormalities were observed in the larvae's body surface, with some larvae failing to complete molting or dying due to an inability to form a normal body surface after molting. After 96 hours, the larval mortality rate was 72.22%, compared to 40% in the GFP control group, demonstrating a significant effect. This invention has broad application prospects in the control of the leaf borer pest in forestry due to its high efficiency, specificity, and safety.

This invention discloses a SUMO-C7H 13 This invention relates to the field of biotechnology, specifically to NO2 probes, their synthesis methods, and applications. The molecular formula of the probe described in this invention is SUMO-C7H. 13 NO2, wherein the SUMO is the human SUMO-1 protein, the nucleotide sequence of which is shown in SEQ ID NO:1 and the amino acid sequence of which is shown in SEQ ID NO:2. This invention obtains the polypeptide sequence encoding the human SUMO-1 protein through gene cloning technology, and further prepares a structurally stable SUMO-C7H protein using chemical synthesis methods. 13 NO2, this probe has a stable molecular structure and is easy to synthesize and detect; SUMO-C7H 13 NO2 fluorescent probes possess high specificity and good detection sensitivity, providing an innovative research tool for exploring tumorigenesis mechanisms and assessing SUMOylation modification levels.

The invention provides an anti-human CD8 rabbit recombinant monoclonal antibody and application thereof. The antibody comprises a heavy chain variable region VH and a light chain variable region VL, a rabbit is immunized with a KLH-coupled CD8 polypeptide antigen, spleen lymphocytes are separated after titer detection is qualified, and single B cell sorting is carried out through a biotin-labeled antigen; positive clone is obtained by a rabbit single B cell culture technology, RNA (Ribonucleic Acid) is extracted and is reversely transcribed into cDNA (Complementary Deoxyribonucleic Acid), and light and heavy chain variable region genes are amplified and cloned into an expression vector for recombinant expression; and finally, the rabbit recombinant monoclonal antibody with high affinity is obtained. The antibody has the capacity of specifically recognizing CD8 + cells in human peripheral blood, is high in affinity and low in animal origin, is suitable for immunodetection such as flow cytometry and ELISA (enzyme-linked immunosorbent assay), and can be used in the fields of immune cell typing, immune function evaluation, immunotherapy monitoring and the like. The invention also provides a light and heavy chain variable region amino acid sequence of the antibody, and a basis is provided for intellectual property protection and further development of the antibody.

The application discloses a recombinant lactobacillus casei expressing sialyltransferase ST6GALNAC1, and belongs to the field of genetic engineering and microbial technology. The recombinant bacterium is obtained by cloning the ST6GALNAC1 gene into a pPG612 vector and electrotransforming the ST6GALNAC1 gene into lactobacillus casei ATCC393. Experiments show that the engineering bacterium can significantly enhance glycosylation modification of intestinal mucus protein MUC2, improve expression of tight junction proteins (ZO-1, Occludin and Claudin-1), inhibit activation of a p38 / MAPK inflammatory pathway, and regulate intestinal flora structure, thereby effectively relieving intestinal injury and systemic inflammatory response caused by salmonella. The application provides a strain basis and technical support for development of a new type of oral microecological preparation.

This invention relates to the preparation and application of a recombinant bivalent vaccine for porcine reproductive and respiratory syndrome (PRRS). The recombinant bivalent PRRS vaccine is prepared using gene recombination technology. The genes encoding the fusion proteins of the major structural proteins GP2, GP3, GP4, GP5, and M protein B-cell epitopes of PRRS-like strains NADC30 and NADC34, as well as their recombinant strains, are cloned into a vector, transformed into host bacteria, and prepared through processes such as fermentation, purification, mixing, and emulsification. Animal experiments show that the recombinant bivalent PRRS vaccine can effectively improve the humoral and cellular immune responses in target animals.

The application discloses application of a pear transcription factor PbbHLH164 in promoting fruit ripening, screens a transcription factor PbbHLH164 which is separated from pear fruits and has the function of promoting fruit ripening, and the nucleotide sequence of the transcription factor PbbHLH164 is shown in the sequence table SEQ ID NO.1. The gene is transiently expressed in pear fruits and stably expressed in pear pulp callus through genetic transformation mediated by agrobacterium, and the result shows that the gene expression amount of ethylene synthesis enzyme and the content of ethylene significantly increase after overexpression of PbbHLH164, and the ripening of the fruits is promoted. Through biological function verification, it is shown that the PbbHLH164 gene cloned in the application has the function of promoting ethylene synthesis to participate in fruit ripening. The discovery of the gene provides a new gene resource for molecular breeding of promoting ethylene synthesis, and development and utilization of the resource are favorable to improve the commodity value of pear fruits, prolong the shelf life of the fruits, and are favorable to reduce agricultural cost and realize environment friendliness.

This invention discloses a dragonfly pineapple AfATL6 This research, encompassing gene cloning methods, expression vectors, and their applications, falls under the field of biotechnology. Specifically, it involves extracting total RNA from dragonfly pineapple, reverse transcribing it to synthesize cDNA, using this as a template to design specific primers, and then employing PCR to obtain... AfATL6 The CDS of the gene was ligated into the pEASY-blunt vector, transformed into E. coli Trans-T1 competent cells, and positive clones were selected to obtain the Dragonfly Pineapple. AfATL6 The gene, whose full-length CDS is 1242 bp, encodes a protein containing 414 amino acid residues. This invention utilizes transient expression and fluorescence microscopy to detect... AfATL6 The subcellular location of the gene-encoded protein is the cell membrane. Through transgenic technology, it was discovered... AfATL6 Gene overexpression delayed flowering in Arabidopsis thaliana and provided a yeast two-hybrid assay to verify the interaction between the AfATL6 protein and AfGF14i, a key component of the florigen-activating complex. AfATL6 Obtaining the gene laid the foundation for studying the flowering mechanism of dragonfly bromeliads and has important theoretical and practical significance.

The application discloses a kind of dsRNA of Sitobion avenae CYP4C1 gene and application thereof, relate to the technical field of agricultural pest control, the cDNA of Sitobion avenae CYP4C1 gene is cloned in the application, its nucleotide sequence is as shown in SEQ ID NO.1;And the dsRNA of the gene is prepared, nucleotide sequence is as shown in SEQ ID NO.2;The preparation method of the dsRNA includes total RNA extraction, cDNA synthesis, target gene cloning, T7 promoter fragment amplification and in vitro transcription purification steps;The dsRNA of Sitobion avenae CYP4C1 gene and application thereof, by feeding method, the dsRNA is introduced into the 3rd instar nymph of Sitobion avenae, and after feeding 48h, CYP4C1 gene silencing efficiency reaches 58%, can significantly reduce the survival rate and the amount of aphid of Sitobion avenae on imidacloprid seed dressing wheat seedling, target strong, prevention and control effect is remarkable, no chemical residue.

This invention belongs to the field of environmental biotechnology, and in particular, it relates to the prothioconazole methyltransferase gene. proS18 Its applications. This invention discovered strains through bioinformatics analysis and gene cloning. Sphingomonas The key enzyme in sp. AJ-1 that degrades prothioconazole is prothioconazole methyltransferase. proS18 The gene was obtained and prothioconazole methyltransferase ProS18 was expressed through an expression vector. HPLC and enzymatic detection confirmed its efficient and specific degradation catalytic ability for prothioconazole, laying the foundation for the development of corresponding biodegradation reagents or efficient degrading bacteria.

This invention belongs to the field of environmental biotechnology, and in particular, it relates to the prothioconazole methyltransferase gene. proS32 Its applications. This invention discovered strains through bioinformatics analysis and gene cloning. Sphingomonas The key enzyme in sp. AJ-1 that degrades prothioconazole is prothioconazole methyltransferase. proS32 The gene was obtained and prothioconazole methyltransferase ProS32 was expressed through an expression vector. HPLC and enzymatic detection confirmed its efficient and specific degradation catalytic ability for prothioconazole, laying the foundation for the development of corresponding biodegradation reagents or efficient degrading bacteria.

This application discloses an epoxide hydrolase gene cloned from Angelica dahurica. AdEH1 and AdEH2 and its encoded proteins and applications, among which, AdEH 1 and AdEH2 The nucleotide sequences are shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively, and the amino acid sequences encoding the proteins are shown in SEQ ID NO: 3 and SEQ ID NO: 4, respectively. AdEH1 and AdEH2 The encoded protein possesses epoxide hydrolase catalytic activity, capable of catalyzing the oxidation of imperatorin and angelica root to produce hydrated oxidized imperatorin and angelica root. This application identifies the gene. AdEH1 and AdEH2 Its key role in the biosynthesis of furanocoumarins provides core gene resources for the regulation and green large-scale synthesis of furanocoumarin components in Angelica dahurica, and also provides technical support for the accurate detection of related compounds.

The application discloses a corn sugar transport gene ZmSTP24 and application thereof. The nucleotide sequence of the corn ZmSTP24 gene is shown as SEQ ID NO. 1, and the amino acid sequence coded by the gene is shown as SEQ ID NO. 2. Through CRISPR / Cas9 technology, the corn ZmSTP24 gene is edited and its function is researched. Compared with a transgenic receptor, the ZmSTP24 transgenic strain has a significantly reduced grain sucrose content, starch content and grain size. It is shown that the ZmSTP24 gene cloned in the application positively regulates the accumulation of grain sugar and other carbohydrates, and influences corn filling and grain size.

This invention discloses a tea tree aldehyde-ketone reductase. CsAKRIVB2 The application of genes in improving plant tolerance to cadmium stress falls under the field of plant genetic engineering. Currently, cadmium-resistant gene resources in tea trees are scarce, limiting the creation of highly resistant varieties. This invention relates to a nucleic acid, the nucleotide sequence of which is SEQ ID NO.1; this invention also relates to… CsAKRIVB2 Genes, Promotion CsAKRIVB2 Reagents for gene expression or CsAKRIVB2 The application of a protein in enhancing plant resistance to cadmium stress, the protein sequence of which is SEQ ID NO.2. This invention clones the gene into the pCAMBIA3101 vector, transforms Arabidopsis thaliana with Agrobacterium GV3101, and obtains transgenic plants. In the Arabidopsis thaliana model, CsAKRIVB2 Overexpression increased the survival rate of transgenic plants by 31.14% and the fresh weight by 131.28% under 25 mM CdCl₂ stress. The molecular mechanism involves activation HMA3 / PCS2 This gene enhances the differentiation, chelation, and induction of SOD / POD / CAT antioxidant enzyme activity. It provides a candidate resource for cadmium-resistant breeding in tea plants.

This invention discloses a method for preparing the BjIM1 protein and gene from amphioxus and its application in lupus erythematosus, belonging to the field of biopharmaceutical technology. The process includes: S1. Material acquisition: collecting amphioxus from Qingdao; S2. Screening and preparation of BjIM1 protein: S2.1, screening for differentially expressed proteins under LPS stimulation; S2.2, screening for differentially expressed proteins with significantly upregulated LPS and low homology to known functional proteins, exhibiting anti-inflammatory activity by inhibiting pro-inflammatory factor expression and regulating immune tolerance; S3. Cloning and preparation of the BjIM1 gene: S3.1, extracting total RNA from amphioxus tissue; S3.2, designing specific primers based on the amino acid sequence of the BjIM1 protein; S3.3, amplifying the gene sequence using RACE technology, and assembling to obtain the full-length BjIM1 gene sequence containing a complete ORF; S3.4, verifying the result by PCR amplification and sequencing, obtaining the amphioxus BjIM1 gene encoding the BjIM1 protein. This invention possesses the technical advantages of definite anti-inflammatory effect, precise immune regulation, high biosafety, no risk of drug resistance, and stable and reproducible process.

The present application relates to the technical field of biometrics and in vitro diagnosis standardization, and particularly relates to a preparation and value determination method of alpha-synuclein solution standard substance, comprising the following steps: cloning a gene coding alpha-synuclein into a prokaryotic expression vector for recombinant expression, purifying the protein raw material with purity greater than or equal to 99% through affinity chromatography and ion exchange chromatography; diluting and sub-packaging the protein raw material for storage to obtain the solution standard substance; determining the value of the standard substance by using an isotope dilution mass spectrometry method based on amino acid analysis, calculating the mass concentration value by measuring the peak area ratio of stable amino acids and isotope-labeled amino acids after hydrolysis, and tracing the value to the international unit system through the amino acid primary standard substance; and performing uniformity testing and stability investigation on the standard substance, solving the problem of lack of standard substance with metrological traceability in the alpha-synuclein detection field, and obtaining the standard substance which can be used for value transmission and detection result standardization among different in vitro diagnosis platforms.

This invention discloses a chitin deacetylase product and its application in agricultural disease prevention. Through codon optimization, the *Rhodococcus* chitin deacetylase gene (SEQ ID NO:1) was cloned into the *Pichia pastoris* expression vector pGAPZa A, constructing the recombinant vector pGAPZaA-CDA-His, which was then transformed into the *Pichia pastoris* SMD1168 host, achieving efficient secretory expression of the enzyme. The resulting recombinant enzyme exhibits high activity and good thermostability. This invention includes a universal enzymatic deacetylation process characterized by deacetylation efficiency, applicable to chitin deacetylases from various sources. This invention also provides a method for further enzymatic hydrolysis to produce N-acetylglucosamine monomers or their oligosaccharides, and the application of the recombinant enzyme and its product in the preparation of biopesticides and the treatment of chitin-containing biomass materials.

The application provides an expression system and application of tilapia antibacterial protein Mptx, and belongs to the technical field of recombinant proteins.The tilapia Mptx eukaryotic protein is obtained by the method of gene cloning and eukaryotic expression vector construction, the binding and agglutination effect of Mptx on related pathogenic bacteria of tilapia are detected by ELISA and agglutination experiment, and it is proved that the obtained Mptx has the effects of inhibiting the proliferation of pathogenic bacteria and protecting the organism by the methods of bacteriostatic experiment and survival rate detection.The application proves that the Mptx eukaryotic protein can significantly reduce the death of tilapia caused by streptococcus agalactiae infection, and has important significance in the aquaculture industry.

The invention provides, inter alia, a nucleic acid (e.g. expression vector) that comprises at least a first coding sequence and a second coding sequence. Each coding sequence is under the control of an inducible promoter of defined strength. Different promoters can have different strengths. Each promoter is responsive to the same inducer. The invention also provides: methods of expressing coding regions, methods of making a product of a multi-enzyme pathway, and methods of optimizing the yield of a product of a multi-enzyme metabolic pathway using the nucleic acids provided by the invention. Also disclosed is a method of non-enzymatic gene cloning useful for practicing the invention.