56 results about "Monokaryon" patented technology

The invention discloses a method utilizing acidic ionic liquid in catalyzing and synthesizing diphenolic acid and / or diphenolic acid ester. The method comprises heating and stirring 1-100 parts of the acidic ionic liquid, 1-200 parts of levulinic acid and 1-500 parts of phenol for 5-60 hours' reaction under the protection of nitrogen to obtain the diphenolic acid; and continuously adding alcohols into the reacted mixture, stirring and heating the mixture to obtain the corresponding diphenolic acid ester. The acidic ionic liquid utilized in the method is sulfonic acid ion liquid including tertiary amine base monokaryon sulfonic acid ion liquid, imidazolyl monokaryon sulfonic acid ion liquid, pyridyl monokaryon sulfonic acid ion liquid, ditertiary amine base dikaryon sulfonic acid ion liquid, diimidazolyl dikaryon sulfonic acid ion liquid or dipyridyl monokaryon sulfonic acid ion liquid. The method successfully enables the acidic ionic liquid to serve as a catalytic agent to prepare the diphenolic acid and / or the diphenolic acid ester and is simple in preparation process and high in yield. According to the method, the separation of the catalytic agent and the reaction substrate is achieved through a solvent extraction method, and the catalytic agent can be repeatedly used and has no significant inactivation.

The invention discloses a rhizoma gastrodiae glucosyltransferase gene. The nucleotide sequence of the rhizoma gastrodiae glucosyltransferase gene is shown as SEQID NO:1, and an amino acid sequence asshown in SEQID NO:2 is coded. A glucosyltransferase gene sequence is screened out according to a CDS sequence provided by a rhizoma gastrodiae transcriptome, a nest type PCR primer is designed, and through amplification and splicing, an overall length sequence of the glucosyltransferase gene is obtained; a prokaryotic expression vector pGEX4T-UGT is constructed, proteins are obtained through expression and purification, a eukaryotic expression vector is constructed, subcellular localization is performed on expression of a UGT gene, glucosyltransferase is transferred into monokaryon honey mushroom mycelia by an agrobacterium tumefaciens infection method for the constructed eukaryotic expression vector pH2GW7.0 35S-UGT, transgene honey mushrooms are obtained, biosynthetic gastrodin is used,the rhizoma gastrodiae is artificially cultured by the transgene honey mushrooms, the content of gastrodin in the rhizoma gastrodiae is increased, and besides, the gastrodin can also be produced through culturing genetic engineering honey mushrooms.

The invention relates to a method for protecting the flammulina velutipes strain by utilizing uracil auxotroph. The method comprises the following steps: hybridizing uracil auxotroph monokaryon NG1-92 of flammulina velutipes with a monokaryon wild type strain of flammulina velutipes, and after fruiting, carrying out single spore isolation, thus obtaining a monokaryon strain; inoculating the monokaryon strain to 5-fluoroorotic acid screening culture medium for culturing, carrying out identification, thus obtaining the uracil auxotroph monokaryon strain, and carrying out mon-mon crossing to obtain a uracil auxotroph dikaryon strain. The method provided by the invention is simple and is low in cost, the strain obtained by adopting the conventional strain separation method can not grow and propagate normally, and thus the flammulina velutipes strain is effectively protected.

The invention discloses a method for identifying the protoplast monokaryon mating types of lepista sordida and a special primer pair SR-4*2 thereof. The method comprises the following steps of: with genome DNAs (deoxyribonucleic acids) of the protoplast monokaryons of two plants of lepista sordida to be identified as templates, performing PCR (Polymerase Chain Reaction) amplification by using a PCR primer pair composed of two single chain DNAs as shown in SEQ ID No.1 and SEQ ID No.2, and detecting the sizes of the obtained PCR products; if the PCR products of the protoplast monokaryons of the two plants of lepista sordida to be identified both contain or do not obtain 500 bp-800 bp of DNA segments, indicating that the mating types of the protoplast monokaryons of the two plants of lepista sordida are the same; and if one plant contains 500 bp-800 bp of DNA segments, while the other plant contains no 500 bp-800 bp of DNA segments, indicating that the mating types of the protoplast monokaryons of the two plants of lepista sordida are different.

The invention discloses a method for identifying a mating type of protoplasted monokaryon of lepista sordida and a special primer pair SR-6*4 thereof. The method comprises the following steps of: by taking the genome DNAs of the protoplasted monokaryons of two plants of lepista sordida as templates, respectively, performing PCR (Polymerase Chain Reaction) amplification by using a PCR primer pair composed of two single-stranded DNAs as shown in SEQ ID No.1 and SEQ ID No.2; detecting the sizes of the PCR products obtained; if the PCR products of the protoplasted monokaryons of the two plants of lepista sordida to be identified both contain DNA segments of 500 bp-800 bp or not, indicating that the mating types of the protoplasted monokaryons of the two plants of lepista sordida to be identified are the same; if one plant contains the DNA segments of 500 bp-800 bp while the other plant contains no DNA segment of 500 bp-800 bp, indicating that the mating types of the protoplasted monokaryons of the two plants of lepista sordida to be identified are different.

The invention discloses a method for identifying the mating types of Lepista sordida protoplast monokaryons and a special primer pair SR-1*10 therefor. The method comprises the following steps: respectively taking genome DNAs (deoxyribonucleic acid) of two Lepista sordida protoplast monokaryons to be identified as templates, performing PCR (polymerase chain reaction) amplification by using a PCR primer pair composed of two single-chain DNAs as shown in SEQ ID NO.1 and SEQ ID NO.2, and detecting the sizes of the obtained PCR products; if both or neither of the PCR products of the two Lepista sordida protoplast monokaryons to be identified contains a DNA segment of 800-1000bp, indicating that the mating types of the two Lepista sordida protoplast monokaryons to be identified are the same; and if one of the two contains a DNA segment of 800-1000bp and the other does not contain a DNA segment of 800-1000bp, indicating that the mating types of the two Lepista sordida protoplast monokaryons to be identified are different.

The invention relates to a method for protecting a lentinus edodes strain through uracil auxotroph.The method comprises the steps that a lentinus edodes uracil auxotroph monokaryon 423-9 and a lentinus edodes monokaryon wild-type strain are crossbred, and single spore isolation is carried out after fruiting to obtain a monokaryon strain; a 5-fluororotic acid screening medium is inoculated with the monokaryon strain for culture, authentication is carried out to obtain a uracil auxotroph monokaryon strain, and then a uracil auxotroph dikaryon strain is obtained by means of mon-mon cross breeding.The method is simple, cost is low, a strain obtained through a conventional strain separation method cannot grow or reproduce normally, and the lentinus edodes strain can be effectively protected through the method.

The invention discloses a method for identifying the mating types of Lepista sordida protoplast monokaryons and a special primer pair SR-1*10 therefor. The method comprises the following steps: respectively taking genome DNAs (deoxyribonucleic acid) of two Lepista sordida protoplast monokaryons to be identified as templates, performing PCR (polymerase chain reaction) amplification by using a PCR primer pair composed of two single-chain DNAs as shown in SEQ ID NO.1 and SEQ ID NO.2, and detecting the sizes of the obtained PCR products; if both or neither of the PCR products of the two Lepista sordida protoplast monokaryons to be identified contains a DNA segment of 800-1000bp, indicating that the mating types of the two Lepista sordida protoplast monokaryons to be identified are the same; and if one of the two contains a DNA segment of 800-1000bp and the other does not contain a DNA segment of 800-1000bp, indicating that the mating types of the two Lepista sordida protoplast monokaryons to be identified are different.

The invention discloses a two-step inoculation method for increasing the fruiting body yield of Cordyceps militaris. The method comprises first inoculating the monokaryotic strain of Cordyceps militaris on the culture medium of Cordyceps militaris and cultivating it for a period of time; Inoculate the Cordyceps militaris monokaryon, heterokaryon or homokaryon strains of the opposite mating type gene of the inoculated monokaryotic strain to the culture of Cordyceps militaris inoculated for the first time, and continue to cultivate until the fruiting body of Cordyceps militaris is harvested . The two-step inoculation method adopted in the present invention can not only reduce the impact of strain degeneration caused by karyotype changes in large-scale production, but also greatly improve the biotransformation rate of the fruiting body of Cordyceps militaris, so the biological yield of the fruiting body of Cordyceps militaris can be effectively improved, The invention is an important innovation in the field of edible fungus strain inoculation.

The invention discloses a method utilizing acidic ionic liquid in catalyzing and synthesizing diphenolic acid and / or diphenolic acid ester. The method comprises heating and stirring 1-100 parts of the acidic ionic liquid, 1-200 parts of levulinic acid and 1-500 parts of phenol for 5-60 hours' reaction under the protection of nitrogen to obtain the diphenolic acid; and continuously adding alcohols into the reacted mixture, stirring and heating the mixture to obtain the corresponding diphenolic acid ester. The acidic ionic liquid utilized in the method is sulfonic acid ion liquid including tertiary amine base monokaryon sulfonic acid ion liquid, imidazolyl monokaryon sulfonic acid ion liquid, pyridyl monokaryon sulfonic acid ion liquid, ditertiary amine base dikaryon sulfonic acid ion liquid, diimidazolyl dikaryon sulfonic acid ion liquid or dipyridyl monokaryon sulfonic acid ion liquid. The method successfully enables the acidic ionic liquid to serve as a catalytic agent to prepare the diphenolic acid and / or the diphenolic acid ester and is simple in preparation process and high in yield. According to the method, the separation of the catalytic agent and the reaction substrate is achieved through a solvent extraction method, and the catalytic agent can be repeatedly used and has no significant inactivation.

The invention discloses a method for identifying the mating types of Lepista sordida protoplast monokaryons and a special primer pair SR-6*6 therefor. The method comprises the following steps: respectively taking genome DNAs (deoxyribonucleic acid) of two Lepista sordida protoplast monokaryons to be identified as templates, performing PCR (polymerase chain reaction) amplification by using a PCR primer pair composed of two single-chain DNAs as shown in SEQ ID NO.1 and SEQ ID NO.2, and detecting the sizes of the obtained PCR products; if both or neither of the PCR products of the two Lepista sordida protoplast monokaryons to be identified contains a DNA segment of 500-800bp, indicating that the mating types of the two Lepista sordida protoplast monokaryons to be identified are the same; and if one of the two contains a DNA segment of 500-800bp and the other does not contain a DNA segment of 500-800bp, indicating that the mating types of the two Lepista sordida protoplast monokaryons to be identified are different.

The invention discloses a method for identifying or assisting in identifying the mating types of Lepista sordid protoplast monokaryons and special primer pairs IS-873 thereof. The method for identifying or assisting in identifying the mating types of the Lepista sordid protoplast monokaryons comprises the following steps: respectively taking the genome DNA of two Lepista sordid protoplast monokaryons to be identified as templates; performing polymerase chain reaction (PCR) amplification on by using PCR primer pairs shown in SEQ ID No. 1 and 2; detecting the size of the obtained PCR products; if the PCR products of the two Lepista sordid protoplast monokaryons to be identified comprise 500 to 600 bp of DNA segments or do not comprise 500 to 600 bp of DNA segments, the mating types of two Lepista sordid protoplast monokaryons are identical; and if the PCR product of one of the two Lepista sordid protoplast monokaryons to be identified comprises 500 to 600 bp of DNA segments and the PCR product of the other one of the two Lepista sordid protoplast monokaryons to be identified does not comprise 500 to 600 bp of DNA segments, the mating types of two Lepista sordid protoplast monokaryons are different.