75 results about "Vitellogenin" patented technology

A universal secretory signal originally derived from a piscine vitellogenin (Vtg) gene is inserted into various expression vectors for driving the secretion of the recombinant protein into the culture medium. This enhances the detection, quantification and downstream scaled-up purification of a recombinant protein of interest. The secretory signal system is very versatile, being conveniently and widely applicable to an array of heterologous host cells such as bacteria, yeast, insect, piscine, and mammalian cell lines (e.g., COS, CHO, NIH / 3T3). The said secretory system is also applicable as a reporter vector for secretion of reporter proteins / enzymes, thus, enabling the detection of the reporter proteins (e.g., CAT, GFP) in the culture medium.

The present invention is directed to a simple method for absolute quantification of plasma vitellogenin from two or more different fish species such as Rainbow trout and Atlantic salmon, or Atlantic cod and haddock. In the case of Rainbow trout and Atlantic salmon, plasma samples obtained from control and β-estradiol induced fish were digested with trypsin. A characteristic ‘signature peptide’ was selected and analyzed by high performance liquid chromatography coupled to an electrospray quadrupole-time-of-flight tandem mass spectrometer, using a deuterated homologue peptide as an internal standard. The hybrid tandem mass spectrometer was operated in a ‘pseudo’ selected reaction monitoring mode by which three diagnostic product ions were monitored for identification and quantification purposes. The reproducibility (coefficient of variation ˜5%) and sensitivity (limit of quantification of 0.009 mg / mL) achieved by this simple assay allow it to be considered as an alternative to immunological assays. In the case of Atlantic cod and haddock, the amino acid sequence of the vitellogenin protein has not yet been determined, but, the Atlantic cod vitellogenin has been characterized using a ‘bottom-up’ mass spectrometric approach. Vitellogenin synthesis was induced ‘in vivo’ with β-Estradiol, and subjected to trypsin digestion for characterization by matrix-assisted laser desorption / ionization-Quadrupole-Time-of-flight tandem mass spectrometry. A peptide mass fingerprint was obtained and ‘de novo’ sequencing of the most abundant tryptic peptides was performed by low energy collision induced dissociation-tandem mass spectrometry. Thus, the sequences of various tryptic peptides have been elucidated. It has also been determined that Atlantic cod vitellogenin shares a series of common peptides with the two different known vitellogenin sequences of Haddock, a closely related species. There are also disclosed novel isolated signature peptides, namely Thr-Tyr-Phe-Ala-Gly-Ala-Ala-Ala-Asp-Val-Leu-Glu-Val-Gly-Val-Arg, Asp Leu Gly Leu Ala Tyr Thr Glu Lys, Phe Phe Gly Gln Glu Ile Ala Asn Ile Asp Lys, Glu Ile Val Leu Leu Gly Tyr Gly Thr Met Ile Ser Lys and Tyr Glu Ser Phe Ala Val Ala Arg.

The invention discloses an enzyme-linked immunosorbent kit for detecting the vitellogenin of goldfish, and a preparation method and applications thereof. The preparation method comprises the following steps: when in preparation, firstly, the pure product of the vitellogenin of the goldfish is prepared; secondly, the polyclonal antibody of rabbit anti-vitellogenin of the goldfish is prepared; thirdly, the polyclonal antibody of the rat anti-vitellogenin of the goldfish is prepared; and fourthly, respectively one bottle of the pure product of the vitellogenin of the goldfish, the polyclonal antibody of the rabbit anti-vitellogenin of the goldfish and the polyclonal antibody of the rat anti-vitellogenin of the goldfish, one block of blank 96-hole enzyme-linked immunosorbent assay board and respectively one bottle of the serum of a negative contrasting group, coating liquid, confining liquid, sample diluent, washing liquid, color development liquid, terminator and goat anti-mouse secondary antibody marked by horse radish peroxidase are filled into a box body together, thus obtaining the enzyme-linked immunosorbent kit for detecting the vitellogenin of the goldfish. The kit of the invention can be applied to the survey that the internal secretion disturbs chemical substances and can quantitatively detect the vitellogenin in the blood of the goldfish, liver tissues and the culture solution of hepatic cells, with sensitivity, accuracy and convenience.

The invention relates to a test kit for qualitatively detecting fish vitellogenin by utilizing lipovitellin antibody. The test kit comprises pure product of goldfish lipovitellin and anti-rat goldfish lipovitellin polyclonal antibody. The preparation method of the test kit comprises the following steps: obtaining goldfish lipovitellin through purifying at first; then preparing anti-rat goldfish lipovitellin polyclonal antiserum through immune rats; and further purifying anti-rat goldfish lipovitellin polyclonal antibody. According to the invention, the test kit can be used for screening environmental estrogen materials and monitoring the situation of water environment which is polluted by the environmental estrogen materials, so that vitellogenin in fish blood, overall uniform serous fluid, skin mucus, liver tissues and hepatic cell nutrient solution can be monitored sensitively and conveniently. The test kit can be used for qualitatively detecting cyprinidae fish vitellogenin, and has the same monitoring sensitivity to the fish vitellogenin of a far provenance relationship fish. The biggest advantage of the test kit is that the test kit has a very wide application scope, and the minimum monitoring limit can reach 100ng / ml.

The invention discloses a kit for detecting that the internal secretion of the sea disturbs chemical substances, and a preparation method and applications thereof. The preparation method comprises the following steps: when in preparation, firstly, the pure product of the vitellogenin of snapper is prepared; secondly, the polyclonal antibody of rabbit anti-vitellogenin of snapper is prepared; and thirdly, one bottle of pure vitellogenin of the snapper, one bottle of the polyclonal antibody of the rabbit anti-vitellogenin of the snapper, one block of blank 96-hole enzyme-linked immunosorbent assay board and respectively one bottle of the serum of a negative contrasting group, coating liquid, confining liquid, sample diluent, washing liquid, color development liquid, terminator and goat anti-mouse secondary antibody marked by horse radish peroxidase are filled into a box body together, thus obtaining the kit for detecting that the internal secretion of the sea disturbs the chemical substances. The kit for detecting that the internal secretion of the sea disturbs the chemical substances can be applied to the detection that the internal secretion disturbs the chemical substances and can quantitatively detect the vitellogenin in the blood of the snapper, liver tissues and the culture solution of hepatic cells, with sensitivity, accuracy and convenience.

The invention relates to a screening method for roes of fishes in Acipenseridae, which aims to solve the problem that the evaluating method is mistaken as the detecting method for the quality of the fish roes is lack of overall evaluation. The evaluation method is as follows, firstly, taking out a roe from the fish in Acipenseridae; secondly, preparing roe extracting solution; thirdly, preparing blood serum; fourthly, preparing body fluid; fifth, determining the polarization indexes of mother cells of the roe; sixthly, determining the density of blood serum vitellogenin; seventhly, determining the contents of Fe ions, roe Zn ions, roe VE and roe glutamic-oxaloacetic transaminase; eighthly, determining the contents of ACP and AKP; and ninthly, if the determined indexes simultaneously meet the target values, the remaining roes in the same ovary are mature, and then the screening of the roes of the fishes in Acipenseridae is accomplished. By adopting the invention, the fertilization rate of the sturgeon, the hatchability of the sturgeon and the survival rate of the fry are more than 85%, 95% and 90% respectively. The screening method is used for breeding the fishes in Acipenseridae.

The invention provides an expression vector for expression of recombinant vitellogenin in an eukaryotic host. An eukaryotic host, including yeast comprising the expression vector according to the invention may be used as a feed or feed additive for both oviparous and non-oviparous animals, including domesticated animals. A transgenic yeast according to the invention contain increased levels of essential amino acids and fatty acids and may be used as a direct feed or fed to an intermediate live feed such as rotifers or artemias to increase the survival rates of oviparous animal or broodstock.

The invention discloses a method for detecting estrogen pollution of water on the basis of polymorphic vitellogenin of periophthalmus modestus, and belongs to the field of molecular biology and environment pollution detection. Amino acid sequences of the vitellogenin of the periophthalmus modestus are shown as SEQ ID NO.2. The method has the advantages that the problems of deficiency of biological in-vivo detection for environmental estrogen by the aid of periophthalmus modestus and deficiency of corresponding biological detection technologies at present can be solved; vivo models of the Periophthalmus modestus which is high in tidal flat settlement property and sensitive to environments are selected, primers are designed, expression of the vitellogenin VtgAa of the periophthalmus modestus is detected by the aid of real-time quantitative fluorescent PCR (polymerase chain reaction) processes, and the method can be used for detecting estrogen pollution degrees of water environments such as seawater and mouths of rivers.

The invention discloses the method for preparing the kit for detecting vitellogenin of cyprinid. The first step is to induce, separate and purify the vitellogenin. The prepared solution of lynoral is injected into the stomach cavity of the cyprinid. With a period of time of induction, the blood serum of the fish blood is obtained through the centrifugal procedure. The freeze-dried powder is prepared by using the separated purification of ion exchange column and desalting of dialysis. The next step is to prepare polyclonal antibody of vitellogenn. The weighted freeze-dried power is taken to dissolve in the double steamed water with Freund's adjuvant being added to emulsify. The polyclonal antibody is prepared through the immune giant blanc. The invention can be used for quantificationally detecting vitallogenin in blood, liver and body of cyprinid with accuracy.

The invention relates to a water estrogen pollution detection method based on oryzias melastigma vitellogenin, and belongs to the field of environmental pollution detection. Currently, no marine fish is used for environment estrogen biological detection, and biological detection technology of seawater estrogen pollution is insufficient. According to the invention, euryhalic male oryzias melastigma with strong adaptability is selected as an in-vivo model; a primer is designed; the expression of vitellogenin (VTG1) is detected by a real-time quantitative PCR method; and 18S ribosome RNA (18S) is used as reference genes to establish a detection system of the VTG1 transcriptional level. Male fish is exposed to 17-beta-estradiol and clean artificial seawater, which are used as a positive and a negative control of the system, and appropriate exposure time is explored. The detection method can be used for detection of environment estrogen pollution degrees of seawater samples, river estuary samples, and saltwater internal lake water samples.

The invention discloses a method for detecting the egg laying amount of insects by a vitellogenin gene, which provides application of a substance which is used for detecting the expression quantity of vitellogenin encoding genes of insect populations eating different substances to detection of the egg laying amount of insect populations eating different substances, wherein the amino acid sequence of vitellogenin is a sequence 2 in the sequence list. Experiments prove that the detection method provided by the invention can be used for detecting the egg laying amount of the insects and particularly the egg laying amount of arma chinensis with different nutrition sources, and the egg laying amount of the insects can be detected within only 2 to 3 days by the detection method while the detection requires 30 to 60 days by a traditional biological method. The detection method provided by the invention has the advantages that materials have wide sources and are easily available, and the operation is simple as well as time-saving and labor-saving, thus being suitable for popularization and application in insect commodity inspection and detection.

The invention relates to breeding for brown planthopper population based on male sterilty after RNAi and a control effect evaluation method. According to the invention, a RNAi breeding method is employed for silencing brown planthopper male testis growth gene PHF7, by detecting the mortality and gene silencing rate, the concentration of dsNlPHF7 is determined; the silenced male insect growth and reproduction parameter indexes are detected, which mainly comprise body weight, life span, accessory gland protein content and arginine content; the copulated male insect growth and reproduction parameter indexes are detected, which mainly comprise body weight, the life span, vitellogenin content, oviposition amount, and YLS richness; and the expression level of the vitellogenin gene in the copulated male insect fat is detected. The brown planthopper population increasing and controlling effects enable quantification evaluation by detecting the important reproduction parameter indexes of the brown planthopper population male insects and female insects, the method has the advantages of simple and fast operation, and has wide applicability, the method provides a novel control approach for preventing and treating brown planthopper, and has theory meaning and utility values.

The invention discloses recombinant yeast for detecting environmental estrogen and a constructing method and use of the recombinant yeast. The recombinant yeast comprises a yeast expression plasmid and a yeast report plasmid, wherein the yeast expression plasmid comprises a tanichthys albonubes estrogen receptor gene TERalpha represented by the sequence shown as SEQ ID No.1, and a carrier of the yeast report plasmid is pMP206, and is linked with a tanichthys albonubes vitellogenin gene promoter 2 represented by a sequence shown as SEQ ID No.4. The tanichthys albonubes estrogen receptor is used for constructing the expression plasmid, the tanichthys albonubes vitellogenin gene promoter is linked with the report gene to construct the report plasmid. The tanichthys albonubes is sensitive to the change of the living environment, and is a wonderful fish biological marker for monitoring the environmental estrogen pollution by using a biological method, and the vitellogenin is a good biological marker for the environmental internal-secretion interfering-substance. The estrogen receptor and the vitellogenin promoter of the tanichthys albonubes are used for constructing a recombinant yeast evaluation system, so that the sensitivity can be greatly improved.

The invention discloses a reagent kit for qualitatively detecting vitellogenin from Pagrosomus major and a preparation method and application thereof. During the preparation, pure vitellogenin from Pagrosomus major is prepared first; then a mouse polyclonal antibody against the vitellogenin from Pagrosomus major is prepared; and then one pure vitellogenin from Pagrosomus major, one mouse polyclonal antibody against the vitellogenin from Pagrosomus major, one PVDF membrane, one chromogenic solution, one confining liquid, one negative control group serum, and one goat anti-mouse secondary antibody labeled by horseradish peroxidase are loaded into a kit body to obtain the reagent kit for qualitatively detecting the vitellogenin from Pagrosomus major. The reagent kit can be applied to the investigation of endocrine disrupting chemicals in a marine environment, and can sensitively, accurately, conveniently and qualitatively detect the vitellogenin in blood, liver tissues, and hepatocyte culture liquid of Pagrosomus major.

The invention discloses a DNA sequence for coding spodoptera exigua vitellogenin (SEVg). The invention also discloses a specific polypeptide sequence capable of utilizing Western blot to detect the AlVg protein expression level. The invention also discloses a polyclonal antibody of an SEVg polypeptide and a preparation method thereof. The polypeptide specific antibody is utilized for analyzing the tissue distribution of SEVg in spodoptera exigua and the difference expression on the protein level. The invention can be used for observing and predicting the population dynamics of spodoptera exigua in fields and can provide material basis and technical support for quick detection and molecular biology study of the protein.

The invention discloses a feed additive capable of promoting the development of ovaries and vitellogenesis of blue crabs and the application of the feed additive. The feed additive is characterized by being prepared through uniformly mixing and compounding the following raw materials in parts by weight: 15-20 parts of DHA ethyl ester oil, 5-10 parts of EPA ethyl ester oil, 10-15 parts of vitamin E acetic ester, 5-10 parts of cholesterol, 5-10 parts of soybean lecithin, 2-5 parts of L-alpha-glycerophosphorylcholine, 1-3 parts of astaxanthin, 1-3 parts of soybean isoflavone, 1-3 parts of lignanoid, 20-25 parts of dried marine algae powder and 10-15 parts of sinonovacula constricta powder. The mass percentage of the additive in released feeds is 0.8-1.0%. The feed additive has the advantages that the survival rate of the blue crabs can be increased by 20-23%, the sexual gland index can be increased by 40-50%, the content of vitellogenin can be increased by 35-45%, the molecule expression quantity of the vitellogenin can be increased by 60-80%, the average weight can be increased by 25-30%, and the breeding yield can be increased by 20-22%.

The invention discloses a method for producing a recombinant rex rabbit nutrient dried meat slice. The method comprises the following steps of: (1) cleaning and picking; (2) smashing; (3) burdening; (4) pickling; (5) pushing on a disk; (6) drying; (7) baking; (8) flattening; and (9) cooling and chopping. According toIn the method, a plurality of nutrient substances such as transglutaminase, soybean protein isolate, ovalbumin, vitellogenin, whey protein, and casein and the like are added into rex rabbit meat, and an organic whole is formed by recombining with rex rabbit meat, so that the original nutrient characteristic of the rex rabbit meat is kept; and a plurality of nutrient elements needed by human bodies are added, so that the nutrient value of the recombinant rex rabbit dried meat slice is raised. The recombinant rex rabbit nutrient dried meat slice produced with the method has particular taste, fragrance, color and luster, good mouthfeel and long shelf life. According toIn the method, the rex rabbit meat of which skin is utilized is taken as a raw material, so that the production cost is lower, and the rex rabbit meat of which the skin is utilized is well utilized comprehensively.

The present invention discloses a vitellogenin peptide fragment tfVWD used for binding to tetrodotoxin. The vitellogenin peptide fragment tfVWD comprises a polypeptide having an amino acid sequence asshown in SEQ ID NO: 1, or an active fragment of the polypeptide. The invention further discloses a nucleotide sequence encoding the vitellogenin peptide fragment tfVWD used for binding to the tetrodotoxin; as shown in SEQ ID NO:2, by taking the vitellogenin peptide fragment tfVWD as an antigen, and mice are immunized to prepare antiserum to obtain a polyclonal antibody. The invention further discloses a preparation method for the vitellogenin peptide fragment tfVWD, and the preparation method comprises the following steps: S1, the nucleotide sequence encoding the vitellogenin peptide fragmenttfVWD is obtained by cloning, and the nucleotide sequence is cloned to a recombinant expression vector; S2, induced expression and condition optimization are performed on the recombinant expression vector to obtain an expression product; and S3, the expression product is filtered, and is eluted through an affinity chromatography column, and elution peaks are collected to obtain recombinant protein. The vitellogenin peptide fragment tfVWD and the nucleotide sequence and polyclonal antibody thereof obtained by the invention have a good binding property with the tetrodotoxin (TTX), and lay a foundation for developing detoxification therapeutic drugs for the tetrodotoxin.

On a surface of a well previously disposed on a plate, a primary antibody that recognizes vitellogenin is solid-phased, in the well a sample obtained from a test body exposed to an environment is injected to react, followed by injecting a secondary antibody that is labeled with an enzyme and recognizes the vitellogenin, further followed by injecting a chromogenic reagent to cause a coloring reaction and by measuring the stained amount, still further followed by calculating an amount of vitellogenin from the stained amount to evaluate an environment based on the amount of vitellogenin.

The invention provides a gobrocypris rarus vitellogenin monoclonal antibody and application thereof. The gobrocypris rarus vitellogenin monoclonal antibody is secreted by a hybridoma cell line with a storing number: CGMCC No.2869. The hybridoma cell line can stably secrete the gobrocypris rarus lvitellogenin monoclonal antibody. A provided kit which comprises the monoclonal antibody can simply, accurately and sensitively detect the vitellogenin in gobrocypris rarus serum in a qualitative / quantitative mode. The Gobrocypris rarus vitellogenin monoclonal antibody is an analysis tool which can simply and effectively detect / evaluate environmental estrogen hormone effect and has wide application prospect.

The invention discloses a tea geometrid vitellogenin gene vtg, which has the nucleotide sequence shown by SEQ ID No.: 1. The invention also discloses a vtg gene encoded protein, which has the amino acid sequence shown by SEQ ID NO: 2. The invention also discloses an application of the vtg gene: the vtg gene can be used to control the reproductive capacity of female Lepidoptera pests.

The invention discloses a DNA sequence used for encoding apolygus lucorum vitellogenin. The invention further discloses a specific peptide sequence capable of detecting an AlVg protein expression level by using Western-blot, and the expression difference of AlVg in apolygus lucorum on the protein level is analyzed by utilizing the specificity of the polypeptide. The invention further constructs an atlas of age in days after female adult eclosion and AlVg expression trend. The correlation of AlVg genes and protein expression levels in female adults with single female fecundity is investigated after the female adults are continuously bred on a variety of host plants, in order to construct a related prediction model, which has significant importance on prediction of the spawning potential of the apolygus lucorum. a positive correlation relation between the AlVg expression levels of female apolygus lucorum adults and the single female fecundity after eclosion for 7 days is defined. The specific peptide sequence disclosed by the invention can be used for observing and predicting the population dynamics of the apolygus lucorum in fields, and providing a foundation for research and development of new technology for monitoring field populations of the apolygus lucorum.

The invention discloses mudskipper's multi-type vitellogenin Vtg gene full-length sequence and a cloning method thereof; amino acid sequences of mudskipper's vitellogenin are shown as in SEQ ID NO. 2, or in SEQ ID NO. 4 or in SEQ ID NO. 6; corresponding genes include VtgAa gene, VtgAb gene and VtgC gene. Type-III Vtg gene full-length CDSs (coding sequences) of mudskipper's Vtg family are acquired herein, the blank in mudskipper's Vtg gene family is filled, and the mudskipper's multi-type vitellogenin Vtg gene full-length sequence and the cloning method thereof are significant to the issues, such as more accurate indication for corresponding water area pollution conditions, and deeper research on the protein expression of aquatics in polluted water areas.

The invention discloses a preparation method of flavored nutritional soft-boiled marinated eggs. The marinated egg process comprises the following steps of cleaning, boiling, cooling, shelling, marination, drying, packaging, sterilization, water washing and cooling, drying and warehousing. The marinated eggs prepared by the method are soft-boiled marinated eggs, and the shelf life of the marinatedeggs can be as long as three months in a normal-temperature storage environment. The marinated eggs are tender and smooth in mouth feel and slightly spicy in taste. Due to the fact that the marinatedeggs are soft-boiled, moisture and nutritional ingredients of the marinated eggs are not lost. The soft-boiled marinated eggs contain rich proteins, fat, vitamins and iron, calcium, potassium and other minerals required by human bodies, are rich in DHA, lecithin and vitellogenin, and contain more vitamin B and other trace elements. The soft-boiled marinated eggs can provide more high-quality proteins, are rich in choline, can improve memory, and can tonify deficiency, benefit vital essence, moisten the lung and tonify the kidney after being eaten for a long time; and rich carbohydrates can supplement glucose consumed by the brain, so that the symptoms of fatigue, irritability, dizziness, insomnia, night sweat, distraction, amnesia and the like caused by insufficient glucose supply of thebrain are relieved.

The invention discloses the method for preparing the kit for fast detecting toxicity of generic estrogen. The first step is to induce, separate and purify the vitellogenin. The prepared solution of lynoral is injected into the stomach cavity of the cyprinid. Then, the freeze-dried powder is prepared by using the separated purification of ion exchange column and desalting of dialysis. The next step is to prepare polyclonal antibody of vitellogenin by using the dissolved freeze-dried powder with Freund's adjuvant being added to emulsify and through the immune giant blanc. The final step is to prepare polyclonal antibody of vitellogenin with the marker of biotin. The invention can be used for quantificationally detecting vitellogenin in blood, liver and body of cyprinid with accuracy.

The invention discloses an important hemiptera natural enemy insect geocoris pallidipennis vitellogenin gene DNA sequence and an encoded protein sequence; hemiptera insect vitellogenin gene molecular characteristic information is complemented, and the new gene is provided for study on an action mechanism of the vitellogenin gene in geocoris pallidipennis nutrition and reproductive regulation and vitellogenin gene functions. A theoretical basis is provided for geocoris pallidipennis mass propagation and effective use, and broad prospects for application in biological prevention and treatment of injurious insects are achieved.

The invention discloses a conopomorpha sinensis bradley vitellogenin gene CsVg, a coding protein and application thereof. The nucleotide sequence of the CsVg gene is shown as SEQ ID NO.1, and the amino acid sequence of the coding protein thereof is shown as SEQ ID NO.2. The invention also discloses a detection method for detecting the influence of the drug on conopomorpha sinensis bradley fecundity, the method detects the expression quantity of the vitellogenin gene CsVg of conopomorpha sinensis Bradley that are applied with and not applied with the drug to judge whether the conopomorpha sinensis bradley fecundity is affected after drug application, has the advantages of high detection sensitivity and accuracy, short detection cycle, good stability, etc., can be used for high-throughput detection of the influence of the drug on conopomorpha sinensis bradley fecundity, and provides molecular reference basis for rational use of the drug in actual production of litchis and longans.

The present invention relates to a microstructure slicing technology, and specifically relates to an acipenser dabryanus oosperm tissue slicing method. The acipenser dabryanus oosperm tissue slicing method mainly comprises seven steps of material fixation, dehydration, transparentizing, wax dipping, embedding, slicing and staining; a fixing liquid used in the material fixation is a Bouin's fixing liquid and a formaldehyde fixing liquid with the mass fraction of 10%; and the total dehydration time is 10-20h. The acipenser dabryanus oosperm tissue slicing method aims at sturgeon oosperm which belongs to a viscid egg, has the characteristics of being rich in vitellogenin, tough and dense in egg envelope, narrow in perivitelline space, and the like, a complete serial tissue slicing method suitable for sturgeon oosperm is established, the sturgeon oosperm can be efficiently processed, and better paraffin sections can be made.