73 results about "Actin genes" patented technology

The invention relates to a kit for gene transcription level of agasicles hygrophila actin ( actin ), and belongs to the technical field of fluorescence-quantitative DNA expansion in vitro in the molecular biological detecting techniques. The invention firstly protects a detecting primer, and the sequence of the detecting primer is showed in SEQ. ID NO. 1 and SEQ. ID NO; secondly the fluorescence-quantitative PCR detection kit capable of detecting the gene transcription level of the agasicles hygrophila actin quantitatively is provided, the operation of the kit is simple, convenient and quick, and the kit is composed of SYBR Premix Ex Taq, primer mixture, a standard actin gene template and ultrapure water. The invention further protects the application, detecting the expresssion quantity of the agasicles hygrophila actin, of the kit. According to the kit for the gene transcription level of the agasicles hygrophila actin, the transcription level of the actin gene can be measured accurately, the high specificity is achieved, the expansion curve shows that the fluorescence signal value of the actin gene is in accord with a standard S-shaped curve, and a melting curve shows that the fluorescent quantitation has high detecting specificity.

The invention relates to a real-time fluorescent quantitative PCR (polymerase chain reaction) kit for one-step quantitative detection of TRECs and KRECs genes and its application. The kit comprises a real-time fluorescent quantitative PCR reaction system based on the real-time fluorescent PCR technique. The real-time fluorescent quantitative PCR reaction system comprises forward and reverse primers specific to TRECs, KRECs and beta-actin genes, and a specific fluorescent probe. The kit allows quick joint screening of neonatal immune system T-cell level and B-cell level, has high sensitivity and stability and provides excellent reproducibility, and this method is applicable to the joint quantitative detection of TRECs and KRECs and to functional screening of the neonatal immune system and is worthy of practical clinical application.

The invention relates to a reference gene for fluorescence quantification of different tissues of Chinese yam and primers and application thereof, and belongs to the technical field of molecular biology of Chinese yam. The nucleotide sequence of the reference gene CKI-2 is as shown in SEQ ID NO. 1. The nucleotide sequence of a PCR amplification primer of the reference gene CKI-2 is as follows: anupstream primer is shown as SEQ ID NO.2; and the downstream primer is as shown in SEQ ID NO. 3. Protein kinase I, myb, F-box, actin gene, EF1, bHLH13, eIF1, grid heavy chain protein and other genes are selected as candidate genes of different tissues. The stability of candidate genes is evaluated through qRT-PCR in combination with RefFind, reference genes stably expressed in different tissues ofthe Chinese yams are selected out through screening, and a reference basis is provided for later development of research work such as gene function verification of growth and development and metabolicmechanisms of the Chinese yams.

The invention discloses a selection method of reference genes in quantitative real-time PCR analysis of jerusalem artichoke, and relates to the field of quantitative PCR. The method comprises the following steps: taking different tissues (radicles, immature stems, leaves, stem blocks and petals) of the jerusalem artichoke as materials, and carrying out expression analysis of the reference genes of18S ribosomal RNA gene (18S rRNA), transcription elongation factor gene (Ef-1a), actin gene (Actin and beta-actin), 3-glyceraldehyde phosphate dehydrogenase (GAPDH), 25S ribosomal RNA gene (25S rRNA)and poly-ubiquitin enzyme gene (UBQ)7 by using a q PCR technology; carrying out statistical assessment on the obtained data and analyzing expression change of all housekeeping genes by utilizing GeNorm and NormFinder software, so as to screen out relatively-stable genes as the reference genes of the jerusalem artichoke, which are used for studying the gene dosage changes of the jerusalem artichoke.

The invention relates to a paeonia ostii reference gene under drought stress, and a special primer and application thereof. The reference gene is a TATA box binding protein TBP gene, an actin ACT1 gene, an actin ACT2 gene, a glyceraldehyde-3-phosphate dehydrogenase GAPDH gene, an eukaryote translation initiation factor eIF1 gene, a eukaryote translation initiation factor eIF2 gene, a tubulin alpha-TUB gene, a tubulin beta-TUB gene, an RNA polymerase II RNA Pol II gene or an RNA polymerase II transcription factor RP II gene. The nucleotide sequence of the gene is disclosed as the sequence table. The invention aims to provide a gene which can be stably expressed in the paeonia ostii gene expression profile under drought stress, and the gene is used as a paeonia ostii drought stress referencegene.

The present invention provides a method and single-tube assay for identification and quantitative analysis of differentially methylated MLH1 promoter sequences that are associated with certain types of cancer in an individual by obtaining a biological sample comprising DNA from the individual, detecting the presence of and measuring the level of methylated MLH1 promoter sequences, and comparing the presence of and level of methylation in the sample to a normalization reference of “normal” beta-actin gene promoters, wherein a difference in the level or pattern of MLH1 methylation of the sample compared to the Actin gene reference level identifies abnormally methylated MLH1 promoter sequences associated with cancer.

A fluorescence quantitative internal reference gene under the drought stress of Haizhou Changshan Mountain, a special primer and the application thereof are disclosed, wherein the fluorescence quantitative internal reference gene is Actin, AP-2 and RAN gene, wherein that gene sequence of the Actin gene is as shown in SEQ ID NO. 1, AP-The gene sequence of RAN gene is shown in SEQ ID NO. 6 and the gene sequence of RAN gene is shown in SEQ ID NO. 13. The invention screens out the candidate gene with stable expression from the transcriptome data and the internal reference gene studied by the predecessor, its stability is evaluated by software, 3 most stable internal reference gene suitable for expression under drought stress in Haizhou Changshan mountain are disclose, Based on these genes, real-time fluorescence quantitative PCR primers were designed, which filled the situation that there were no internal reference genes under drought stress in Haizhou Changshan Mountain, and provided a strong support for the precise quantification of related functional genes under drought stress in Haizhou Changshan Mountain, and could improve the stability, repeatability and reliability of the research.

The present invention aims at providing one homologous recombination method for in-situ fish gene modification breeding. The method includes the following steps: 1) separating specific fish gene flanking sequence; promoter regulating sequence DNA segments to constitute homologous recombination vector with actin gene promoter regulating sequence to replace growth hormone gene promoter regulating sequence; 2) microscopic injecting the homologous recombination vector to zebra fish embryo to screen positive homologous recombination replaced fish fry; and 3) analyzing relative gene expression level and genetic phynotype of the endogenous gene modified fish. The present invention is used in breeding fish variety.

The invention relates to the field of molecular detection, particularly a primer set and probe for apple Actin gene real-time fluorescent quantitative PCR (polymerase chain reaction) detection and a detection method by using the same. The apple Actin gene real-time fluorescent quantitative PCR detection method provided by the invention comprises the following step: carrying out real-time fluorescent quantitative PCR detection on a sample to be detected by using Act-Probe-F and Act-Probe-R as primers and Act-Probe as a probe. The apple Actin gene real-time fluorescent quantitative PCR detection method provided by the invention has the advantages of high specificity, favorable repetitiveness and high sensitivity; the lower detection limit is 1.48*10<1>copies.mul<-1>; and the sensitivity is 1000 times of that of the conventional RT-PCR (reverse transcription-polymerase chain reaction) technique. The invention lays the foundation for quantitative analysis on expression of apple functional genes and pathogenetic genes by using the real-time fluorescent quantitative PCR method.

The invention discloses preparation of a chicken eimeria tenella strain with EYFP knocked into an Et.Actin gene. According to the invention, an FnCas12a protein, crRNA synthesized in vitro and a homologous recombination fragment (an Et.Actin gene containing a P2A-DHFR-EYFP expression cassette) are introduced into chicken eimeria tenella sporozoite by utilizing a Lonza 4D-Nucleofector nuclear transformation instrument, so that the editing of the chicken eimeria tenella is realized. The edited insect strain is screened through pyrimethamine in a chicken body to obtain a high-purity gene edited insect strain. Then, the gene edited insect strain is verified by utilizing PCR and Sanger sequencing technologies, and it is proved that the EYFP successfully marks the Et.Actin gene. After sporozoites of the strain are subjected to cell culture and a chicken is infected, cecum mucosa is taken and smeared to show that the EYFP is expressed in the whole life history of the gene edited strain. The gene editing method can be used for performing fixed-point marking on a chicken coccidiosis gene, and is beneficial to analyzing the function of the coccidiosis gene. The invention also has important significance for research and development of novel anticoccidial drugs and vaccines.

The invention provides a vector for expressing and transferring gene with muscle tissue specific expression of phosphoenolpyruvatecarboxykinase (PEPCK) for reducing hyperlipidemia, delaying aging, andimproving reproductive capacity. The vector contains a reconstructed promoter of human alpha-skeletal actin gene, an intron of minute virus of mice (MVM), reconstructed cDNA of human phosphoenolpyruvatecarboxykinase 1 (PCK1), and four human miR-122 target sequences which are connected in series connection. The recombinant vector is mediated by an adeno-associated virus vector, and comprises a type 1 adeno-associated virus and the like.

The invention provides a combination of specific primers and probes and a kit for fluorescent quantitative PCR detection of type 15 HPV. 3 upstream and 2 downstream primer sets and 15 specific Taqman probe sets designed for the L1 region of the HPV gene in the present invention, the corresponding kits can be quickly and easily detected in the same tube under specific PCR conditions, including16,18 , 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, 68, 73 and other 15 high-risk HPV types and 16, 18 HPV types, reduce the cost, and do not need to open The lid operation prevents contamination, and the human actin gene is used as the internal standard gene to reduce the occurrence of false negatives.

The invention relates to a real-time fluorescent quantitative PCR (polymerase chain reaction) kit for one-step quantitative detection of KRECs gene and its application. The kit comprises a real-time fluorescent quantitative PCR reaction system based on real-time fluorescent PCR technique. The fluorescent PCR reaction system comprises forward and reverse primers specific to KRECs and beta-actin genes, and a specific fluorescent probe. The kit allows quick screening of neonatal immune system B-cell level, has high sensitivity and stability and provides excellent reproducibility, and this method is applicable to the quantitative detection of KRECs and to the functional screening of the neonatal immune system and is worthy of practical clinical application.

The invention relates to a real-time fluorescent quantitative PCR (polymerase chain reaction) kit for one-step quantitative detection of TRECs gene and its application. The kit comprises a real-time fluorescent quantitative PCR reaction system based on real-time fluorescent PCR technique. The fluorescent PCR reaction system comprises forward and reverse primers specific to TRECs and beta-actin genes, and a specific fluorescent probe. The kit allows quick screening of neonatal immune system T-cell level, has high sensitivity and stability and provides excellent reproducibility, and this method is applicable to the quantitative detection of TRECs and to the functional screening of the neonatal immune system and is worthy of practical clinical application.

The invention relates to an MTHFR gene polymorphism detection primer. The MTHFR gene polymorphism detection primer comprises a forward primer used when an MTHFR gene c.1298 locus is in an A condition, a reverse primer used when the MTHFR gene c.1298 locus is in the A condition, a forward primer used when the MTHFR gene c.1298 locus is in a C condition, a reverse primer used when the MTHFR gene c.1298 locus is in the C condition, a forward primer used when an MTHFR gene c.677 locus is in the C condition, a reverse primer used when the MTHFR gene c.677 locus is in the C condition, a forward primer used when an MTHFR gene c.677 locus is in the T condition, a reverse primer used when the MTHFR gene c.677 locus is in the T condition, a forward primer for detecting a beta-actin gene and a reverse primer for detecting the beta-actin gene. An MTHFR gene polymorphism detection kit is rapid and convenient to detect, high in sensitivity, high in accuracy rate and low in cost.

The invention provides a method for quantitatively detecting colletotrichum gloeosporioides by utilizing real-time fluorescent PCR (Polymerase Chain Reaction). A real-time fluorescent quantitative PCR detection method of colletotrichum gloeosporioides is established through primers designed by actin genes of colletotrichum gloeosporioides; the detection method disclosed by the invention is high in specificity and sensitivity, can detect colletotrichum gloeosporioides in an incubation period, and has important significance for detecting potential colletotrichum gloeosporioides infection; according to the detection method disclosed by the invention, the normalization relation between pathogenic bacterium DNA and plant DNA biomass is also considered, the growth conditions of colletotrichum gloeosporioides in different host plants can be accurately determined, the method can be used for screening and identifying disease-resistant varieties of arabidopsis thaliana and stylosanthes guieosporioides, and a basis is provided for early detection and timely prevention and treatment of colletotrichum gloeosporioides.

The present invention is directed to the isolation and identification of a non-graminaceous monocotyledonous plant promoter, the banana (Musa.Spp.) actinn gene associated promoter region. The promoter region has been found to unexpectedly direct constitutive gene expression not only in non-graminaceous monocotyledonous plants but also in graminaceous monocotydonous plants. The Invention is also concerned with a chimeric nucleic acid construct comprising the promoter of the invention oprably linked to a foreign or endogenous polynucleotide encoding a protein of interest or a transcript capable of modulating expression of a target gene. The invention further discloses transformed plant cells, as well as differentiated plants and plant parts, containing the construct.

The invention discloses a primer for judging bombyx mori moth formation time. Actin gene 3 and cocoon lyase gene of bombyx mori chrysalis to be detected are taken as templates, and real-time fluorescent quantitative PCR reaction is performed by using the primer to obtain transcription expression quantity of the actin gene 3 and transcription expression quantity of the cocoon lyase gene separately;the relative transcriptional expression quantity of the cocoon lyase gene is the transcriptional expression quantity of the cocoon lyase gene / the transcriptional expression quantity of the actin gene3; the relative transcriptional expression quantity of the cocoon lyase gene is substituted into a correlation model of the relative transcriptional expression quantity of the bombyx mori chrysalis cocoon lyase gene and pupal stage development time; and the bombyx mori moth formation time is judged. According to the method, the pupal stage development time of one or more bombyx mori chrysalis canbe accurately measured to accurately judge the bombyx mori moth formation time, errors caused by traditional naked eye observation can be reduced, and a new detection method is provided for judging the moth formation time in bombyx mori seed production work.

The invention discloses a plant expression vector containing glyphosate resistant gene. The plant expression vector is an annular carrier containing two copies of glyphosate resistant gene; the glyphosate resistant gene coded amino acid sequence is the protein shown as SEQ ID NO.1 or glyphosate resistant protein that is substituted and / or deleted and / or added with one or several amino acid residues compared with SEQ ID NO.1. The two copies of glyphosate resistant gene are respectively driven by a 1, 5-ribulose bisphosphate carboxylase gene promoter from cotton and a promoter from arabidopsis thaliana actin gene. The invention proves that transgenic cotton introducing the plant expression vector has very strong glyphosate resistance. The plant expression vector provided by the invention is of great significance for further cultivation of high glyphosate resistant plants and application thereof.

The invention relates to an RT-RIA / CRISPR-Cas12a detection kit for an The kit comprises a primer and a probe sequence, wherein the primer and the probe sequence are used for detecting RT-RIA / CRISPR-Cas12a of the Asian type Zika virus. The sequences of the primer and the probe comprise RIA primer sequences of Zika virus ZIKV and a reference gene beta-actin gene, a Cas12a reaction crRNA sequence and a fluorescence report probe sequence. And the 5'end of the crRNA sequence contains a T7 promoter recognition site. The fluorescent group marked at the 5' end of the ZIKV and beta-actin gene fluorescent report probe sequence is one of FAM, HEX, ROX, VIC and CY5. And a quenching group marked at the 3'end of the ZIKV and beta-actin gene fluorescence report probe sequence is one of TAMRA, MGB, BHQ1 and BHQ2. The fluorescent groups marked at the 5'ends of the probe ZIKV and the beta-actin are different, the kit further comprises an RIA reaction reagent component, a Cas12a reaction reagent component and a sample releasing agent, the RIA reaction reagent component comprises an RIA Buffer, an RIA enzyme and an RIA activating agent, and the Cas12a reaction reagent component comprises a Cas Buffer and a Cas12a enzyme.

The invention discloses a herbicide resistance gene expression vector and application thereof. The expression vector comprises a promoter, a glyphosate resistance gene, a glufosinate-ammonium resistance gene and a terminator, wherein the glyphosate resistance gene adopts a promoter pUbi, the glufosinate-ammonium resistance gene adopts a composite promoter, and the composite promoter is composed of a corn Ubi promoter pUbi, a 35S promoter of cauliflower virus and an intron in a rice Actin gene. According to the invention, the glyphosate resistance gene and the glufosinate-ammonium resistance gene are highly expressed by using the same vector, and a transgenic plant with high resistance to glyphosate and glufosinate-ammonium at the same time can be obtained through the vector.

The invention discloses a kit and a method for visually detecting trichomonad vaginalis based on an RPA-CRISPR-Cas12a system, the kit comprises a pair of specific RPA primer pairs for amplifying actin genes of trichomonad vaginalis, crRNA for CRISPRCas12a reaction, a fluorescent reporter group, a lateral flow immunochromatography reporter group and a Cas12 / 13 universal lateral flow chromatography test strip, the buffer solution and the Cas12a protein are used for CRISPR (clustered regularly interspaced short palindromic repeats) reaction, nucleotide sequences of a specific RPA (recombinase polymerase amplification) primer pair for amplifying actin genes of trichomonad vaginalis are shown as SEQ ID NO: 11 and 16, and a nucleotide sequence of DNA (deoxyribonucleic acid) complementary with crRNA (complementary ribosomal ribonucleic acid) for CRISPR Cas12a reaction is shown as SEQ ID NO: 5; the method is rapid, visible, good in specificity and high in sensitivity, can be used as a tool for clinical detection of trichomonad vaginalis, has low requirements on equipment, is particularly suitable for large-scale population screening, and provides a good technical means for monitoring of public health institutions.

The invention discloses a kit and method for quantitatively detecting the methylation degree of a human MGMT gene. The kit comprises an MGMT gene detection reagent, an internal control beta-Actin genedetection reagent and a standard substance, wherein the MGMT gene detection reagent comprises an MGMT primer pair and an MGMT probe, and the sequences of the MGMT primer pair and the MGMT probe are as shown in SEQ ID No. 1 to SEQ ID No. 3 respectively; the internal control beta-Actin gene detection reagent comprises a beta-Actin primer pair and a beta-Actin probe, and the sequences of the beta-Actin primer pair and the beta-Actin probe are as shown in SEQ ID No. 4 to SEQ ID No. 6 respectively; the two probes are modified by different fluorophores; and the standard substance is formed by mixing a 100% methylated MGMT gene and a 0% methylated MGMT gene according to different proportions. According to the kit, dual real-time fluorescent quantitative PCR is adopted, a standard curve is drawnby utilizing the standard substance, the methylation degree of the MGMT gene is relatively quantified, and the kit is simple and convenient to use and low in cost.

The invention discloses internal reference genes suitable for researching the flooding stress gene expression of lotus. The internal reference genes include a lotus actin gene and a ribosomal protein 18sRNA gene. The invention discloses a real-time fluorescent quantitative PCR primer designed based on the two genes to solve the problem of no internal reference genes in the researches of the flooding stress gene expression of lotus. The internal reference genes can stably express during the flooding stress of the lotus, and the designed real-time fluorescent quantitative PCR primer can be used for the gene expression analysis of the lotus responding to the flooding stress in order to improve the stability, the reliability and the repeatability of the researches.