57 results about "Vitellogenins" 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 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 present invention belongs to the field of life science. Said invention provides a regulatory controlling sequence expressed by zebra fish vitellogenin 1, (vtg 1) gene of sequence 1. Said invention utilizes the transgenic zebra fish experiment to show that said sequence has promoter activity in the zebra fish body interior, and can make downstream gene implement specific expression in liver under the induction of female sex hormone. By adopting the connection of zfvtg 1-1.7 sequence provided by said invention and reporter gene (fluorescin) the fluorescence can be observed in the living body interior of transgenic zebra fish so as to quickly and visually reflect expression of vtg 1.

The invention discloses a method for regulating and controlling a vitellogenin level of zebra fishes. The method comprises the steps of: quantifying content of vitellogenin in blood plasma, whole body homogenate or head-tail homogenate of an aquatic organism to be detected; when the content of the vitellogenin in the blood plasma, the whole body homogenate or the head-tail homogenate of the aquatic organism to be detected is higher than the content of the vitellogenin in the blood plasma, the whole body homogenate or the head-tail homogenate of a reference aquatic organism, indicating that the aquatic organism to be detected is poisoned or interfered by a low-concentration estrogen interferent. The method disclosed by the invention has the advantages of being capable of detecting whether the aquatic organism is poisoned or interfered by median- and low-concentration estrogen interferents in an exogenous environment and having important implications for developing relevant scientific research, environment pollutant supervisory control, safety management and the like.

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 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 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 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 method for regulating and controlling the mRNA (messenger Ribonucleic Acid) level of zebra fish vitellogenin. The method is used for checking the toxicity or interference for aquatic organisms caused by low-concentration estrogen disruptors. The method comprises the following steps of: firstly, respectively extracting total RNA of livers of the aquatic organisms to be detected and a control aquatic organism; secondly, quantifying the mRNA expression level of VTG1 (Vitellogenin 1) and/or VTG3 of the vitellogenin of the extracted total RNA of the livers; and thirdly, if the mRNA expression level of the VTG1 and/or VTG3 of the vitellogenin of a sample to be detected is higher than the expression level of the VTG1mRNA or VTG3mRNA of the vitellogenin of a control sample, showing that the endocrine secretion in the aquatic organisms to be detected is poisoned or interfered by the low-concentration estrogen disruptors. According to the method disclosed by the invention, whether the aquatic organisms are poisoned or interfered by the low-concentration estrogen disruptors in an external environment or not can be accurately and sensitively detected; and the method has the advantages of favorable sensitivity, high accuracy and the like and can be applied to the field of detection of estrogen disruptors for the aquatic organisms.

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 invention discloses application of a scatophagus argus vitellogenin gene to detection of water area environmental hormones. The application comprises the following steps: obtaining a VTGAb full-length sequence according to a RACE method; sampling and extracting liver RNA (Ribonucleic Acid) after injecting estrogen into scatophagus argus in a gradient manner; detecting expression of VTGAb by adopting an RT real-time quantitative PCR (Polymerase Chain Reaction) method; taking EF1-a as an internal control gene and carrying out absolute quantification on cDNA (complementary Deoxyribonucleic Acid) of liver tissues to establish a detection system of a VTGAb transcriptional level; putting scatophagus argus which is not sexually mature into a water body to be detected and sampling and extracting the liver RNA; detecting expression amount of the VTGAb; and comparing with the detection system, and analyzing the pollution degree of the environmental hormones of a water sample. By adopting the application disclosed by the invention, the pollution condition of the environmental hormones can be measured under the condition that the economic loss is relatively small and the single variable of the scatophagus argus is controlled; and the scatophagus argus vitellogenin gene can be used for the pollution degree of the environmental hormones of water samples of seawater, the mouth of a river and a saline water internal lake.

The invention discloses a fat greenling vitellogenin sandwich ELISA kit and a preparation method, a detection method and application thereof. The preparation method comprises the following steps: obtaining vitellogenin from 17 (i) beta (/ i) - estradiol induced fat greenling plasma through an ammonium sulfate precipitation and SephadexG-200 by purification to prepare a polyclonal antibody, and labeling a horseradish peroxidase enzyme to the purified antibody; putting a fat greenling vitellogenin purifying product, a rabbit anti- fat greenling vitellogenin polyclonal antibody, a horseradish peroxidase enzyme labeled rabbit anti-fat greenling vitellogenin polyclonal antibody, and a blank 96-well ELISA plate as well as a coating buffer, a scrubbing solution, a confining liquid, a sample diluent, a developing solution and a stop buffer in a kit body to obtain a sandwich ELISA kit for detecting the fat greenling vitellogenin. Through the kit provided by the invention, vitellogenins in fat greenling plasma, body surface mucus, a liver tissue and a liver cell culture medium can be sensitively, accurately and quantitatively detected in a detection range of 1.95 to 125ngmL<-1>, and an important tool is provided for the detection of an endocrine disruptor in an offshore area in China.

The present invention relates to the control of pest infestation by inhibiting or reducing the expression of genes of the family of chitin synthase and of vitellogenin, as well as through the expression of the toxin Cry8ka5. The invention further provides method and compositions for controlling pests, by feeding the pest with one or more double-stranded RNA molecules provided by the present invention, as well as double-stranded RNA molecule provided by the present invention, as well as through the action of the toxin Cry8ka5 on the target insect. The invention further describes a method of obtaining transgenic plants that express double-stranded RNA molecules and the toxin protein Cry8ka5. The present invention is preferably used for cotton plants.