33 results about "Pratylenchus" patented technology

The invention provides the identification of the presence of mutant ROS protein in human cancer. In some embodiments, the mutant ROS are FIG-ROS fusion proteins comprising part of the FIG protein fused to the kinase domain of the ROS kinase. In some embodiments, the mutant ROS is the overexpression of wild-type ROS in cancerous tissues (or tissues suspected of being cancerous) where, in normal tissue of that same tissue type, ROS is not expressed or is expressed at lower levels. The mutant ROS proteins of the invention are anticipated to drive the proliferation and survival of a subgroup of human cancers, particularly in cancers of the liver (including bile duct), pancreas, kidney, and testes. The invention therefore provides, in part, isolated polynucleotides and vectors encoding the disclosed mutant ROS polypeptides (e.g., a FIG-ROS(S) fusion polypeptide), probes for detecting it, isolated mutant polypeptides, recombinant polypeptides, and reagents for detecting the fusion and truncated polypeptides. The identification of the mutant ROS polypeptides enables new methods for determining the presence of these mutant ROS polypeptides in a biological sample, methods for screening for compounds that inhibit the proteins, and methods for inhibiting the progression of a cancer characterized by the mutant polynucleotides or polypeptides, which are also provided by the invention.

Relating to the quarantine inspection field of plant diseases and insect pests, the invention provides a Pratylenchus penetrans detection kit and a detection method thereof. By designing specific primers specific to the 18S specific sequence of Pratylenchus penetrans and applying a PCR amplification technology, rapid detection of Pratylenchus penetrans is realized. With the characteristics of strong specificity, high sensitivity and good stability, the method plays an important role in improving the accuracy of Pratylenchus penetrans detection and shortening the detection time.

The invention discloses a natural passage cell line for lamb testis sertoli cells and application of the natural passage cell line to separate culture and proliferation of goatpox viruses. The cell line disclosed by the invention is a lamb testis sertoli natural passage cell line which is obtained by separating and purifying primary cells of a lamb testis to obtain the lamb testis sertoli cells and performing natural passage on the lamb testis sertoli cells. A research shows that the cell line is relatively sensitive to inoculation of the goatpox viruses; the virus titer proliferated by the cell line is greater than that of the primary cells, Vero cells and BHK21 cells of the testis of a newborn lamb by about 1.5, 2.5 and 2.3 respectively. The cell line for separating the goatpox viruses can guarantee the uniformity and the stability of the viruses and prevent the risk that the separated viruses carry other pathogenies due to the fact that the primary cells are used for multiple times. Furthermore, by the use of the passage cell line for preparing vaccines, the production process is simplified, the production period is shortened, and the production cost is reduced; meanwhile, the quality of the vaccines are kept stable; the cell line has certain practical significance for large-scale production of the vaccines.

The invention discloses a loop-mediated isothermal amplification (LAMP) primer group for rapidly detecting pratylenchus thoreni. The primer group comprises an outer primer pair ptF31/ptB31 and an inner primer pair ptFIP1/ptBIP1, wherein the sequences of the primers are respectively shown in SEQ ID NO.1-4. An LAMP method is established based on the primer group and is characterized by using deoxyribonucleic acid (DNA) of samples as a template to carry out LAMP. The results can be judged by the following two methods after reaction ends: one method is that amplification products undergo electrophoresis and the samples with specific ladders are judged to be positive; the other method is that the samples having color change observed with the naked eye are positive by adding SYBR green I to a reaction system. The LAMP method has low requirements for instruments and equipment, is rapid and safe, has good specificity and strong sensitivity, provides technical support for detection of pratylenchus thoreni, especially pratylenchus thoreni rapid detection work of grassroots quarantine units, and has good popularization and application values.

The present invention relates the function of Dickkopf 3 (Dkk3 ) in testis using shRNA mediated knock down model. Specifically, the present invention provides knock down model comprising reduction in Dkk3 activity. The knock down model of Dkk3 consisting of non human vertebrates which have, incorporated in their genome, shRNA construct targeting mammalian Dkk3 gene exhibits low testis weight, low sperm count and has low litter size. The knock down model displays disrupted seminiferous tubules and are subfertile. The present invention model has a role in sex determination as its interruption leads to sex reversal of XY gonads, converting males to females. Sex reversal role of Dkk3 knock down model can find its utility in agricultural applications. The present invention describes Dkk3 as a dual function protein which is associated with sex determination as well as is essential for the process of spermatogenesis. Such testicular functional studies are useful as a model for various disease states of infertility or subfertility and for identifying a potential treatment to overcome idiopathic infertility.

The present invention discloses a porcine parvovirus strain, which is named Parvoviridae, and has the preservation number of CCTCC NO:V201313. According to the present invention, a PCR method is adopted to carry out PCR detection on suspected-PPV disease materials collected in the Henan region, inoculation on porcine testis (ST) cells is performed, isolation is performed to obtain a strain of virus named HP104, and results of cytopathic effect observation, biological characteristic research, virus shape observation through electron microscopy and other approaches confirm that the isolated virus is the porcine parvovirus; and the porcine parvovirus HP104 virus solution and an oil emulsion are mixed to prepare an emulsifier, the emulsifier is inoculated on health guinea pigs, a HI antibody level achieves 128 after 28 days, and porcine parvovirus antibody in the control group guinea pigs is negative, such that the results show that the Henan isolated strain PPVHP104 has good immunogenicity.

The invention discloses a method for rapidly breeding ST (swine testis) sensitive cell lines of a hog cholera lapinized virus strain (C strain) by virtue of a reverse transcription PCR (RT-PCR) (reverse transcription-polymerase chain reaction) technology and a real-time fluorescent quantitative RT-PCR technology. The method comprises the following steps: 1) acquiring ST monoclonal cells; 2) inoculating the hog cholera lapinized virus strain (C strain); 3) extracting RNAs of various batches of hog cholera lapinized virus liquid (virus liquid) which are continuously harvested, and determining that ST monoclonal cell lines can continuously infect the hog cholera lapinized virus strain (C strain) by virtue of the RT-PCR technology; and 4) determining that the hog cholera lapinized virus strain of which the number of copies is 10<6> copies/mL or above are the ST sensitive cell lines of the hog cholera lapinized virus strain (C strain) by virtue of the real-time fluorescent quantitative RT-PCR technology. With the application of the method disclosed by the invention, the sensitive cell lines can be rapidly provided for the production of hog cholera lapinized virus vaccines, so that a vaccine production cycle is shortened and vaccine cost is reduced.

The invention relates to the technical field of livestock breeding, in particular to a breeding method of improving reproductive performance of Angus bulls. The breeding method includes: on the basis of daily breeding management, feeding calves 7 days after weaning with a zinc-contained preparation, and well regulating secretion of gonad activity and sex hormone of the bulls. The zinc-contained preparation is fed after weaning and before mating, modes of massage, acupuncture and the like are adopted, and dosage, drug delivery way, number of times, preparation content and daily feed nutrients are controlled, so that zinc element is sufficiently absorbed by each part of bodies, absorption and digestion are promoted, and metabolism level of multiple hormones in the bodies of male breasts is regulated; meanwhile, absorption conversion of the bodies on the feed is promoted, content of adverse hormones in the bodies is reduced, sperm density and sperm activity are improved, rate of sperm deformation is reduced, and growing development of testis is promoted; liver steroid metabolism is further improved, and sex steroid receptor level is regulated.

The invention discloses a breeding method for a pig frog. The method comprises the following steps of: selecting more than one pure and sexually mature male pig frog to paunch to take the testis of the pig frog out; holding in a vessel with normal saline solution; picking the involucra of the testis off by using a tool; and breaking the testis by clamping and releasing sperms to prepare sperm suspension. According to the method, since the sperms and ova of parent pig frogs of different varieties are selected, and outbreeding is performed by using an artificial fertilization method, the artificial reproduction method of hybridizing the pig frogs breaks through the conventional reproduction mode through natural spawning of the pig frogs and has great realistic significance and historical significance on promoting artificial breeding of special economic animals and particularly the pig frogs so as to improve the yield and the quality of the commercial pig frogs, enriching the vegetable basket of people, accelerating the pace of banishing poverty and becoming prosperous for rural population and even guaranteeing of sustainable and stable development of national economy.

The invention discloses a purification method of recombinant viruses with fluorescent markers. The purification method comprises the following steps: 1, goat pox viruses with KLP2 genes being lost arerecombined, specifically, (1) sheep testicular primary cells are cultured, a cell plate is spread, after cell culture reaches 80% or so, the cells are infected through goat pox virus vaccine strains,and (2) virus recombination is conducted; and 2, the recombinant viruses are purified, specifically, (1) the recombinant viruses with fluorescence are picked, (2) the viruses are purified, (3) purification is continued, specifically, the viruses, with the fluorescence, picked in the previous step are repeatedly subjected to freeze thawing, then according to operation of (2) in the step 2, dilution and inoculation are conducted, then picking is conducted, and generally after 3-4 generations, the purified recombinant viruses can be obtained.

The present invention provides a method of culturing PAMAM-D mediated sperm vector of transgenic mammalian, including following steps: foreign gene enzyme is cut into a linear configuration of end cohesive, determining its dosage range aimed at different sperms; screening PAMAM-D in the composition algebra which is more than G5 and ranges of exogenous charge ratio 2-50:1, transfecting sperm by the exogenous DNA has high effect, and also keeping the optimum combination condition of the exogenous DNA usage of sperm activity and PAMAM-D generation and dose, so as to prepare PAMAM-D/exogenous DNA compound, adequately washing collected sperm under the monitor of the supernatant fluid IF-1 after washed; transfecting sperm outside of compound by incubates method, the female animal is conceived by individually selecting single sperm of transfection exogenous DNA, or sperm of transfection exogenous DNA formed by injecting in testicle, in vitro fertilization embryo transfer, in vivo artificial insemination and natural mating, cultivating transgenic animal.

A detection method for 305 fertility-associated sperm localization protein expressed in human testis and epididymis. The method includes qualitatively or quantitatively detecting for sperm localization proteins on a sperm or in seminal plasma via capture agents for the sperm localization proteins. The sperm localization proteins belong to an important protein group related to human sperm maturation and fertility. The method provides a diagnostic approach for male infertility.

The invention relates to a combined inactivate vaccine for sheep pox and contagious caprine pleuropneumonia (CCPP), and a production method of the combined inactivate vaccine. The method comprises thesteps of inoculating suspension cultured BHK-21 cells and a proper culture medium with a capripox virus AV41 strain and a mycoplasma capricolum pneumonia subspecies C87001 strain respectively for culturing, harvesting the culture, concentrating and purifying; inactivating with BEI and thimerosal respectively, adding a suitable adjuvant according to a certain proportion, mixing and emulsifying. The combined inactivate vaccine is used for preventing the sheep pox and the CCPP. The combined inactivate vaccine, provided by the invention, for the sheep pox and the CCPP is obtained internationallyfor the first time, and can achieve the aim of preventing the two diseases by means of one injection; furthermore, a live vaccine for the sheep pox is produced by means of the suspension cultured BHK-21 cells; compared with the original inactivate vaccine prepared from ovine testicular cells, the combined inactivate vaccine provided by the invention has obvious advantages, and can solve the problems that in the production process of the live vaccine for the sheep pox, the sheet testis is difficult to obtain, lamb death and ewe abortion are easily caused, subcutaneous immunization needs to be performed when the vaccine is used, an operation is difficult during clinical application, and the like.

The invention relates to a constructed shRNA lentiviral interference vector with the optimal efficiency for suppressing the expression of chicken cyclin F genes. Four shRNA sequences designed for the chicken cyclin F genes are copied into a lentiviral vector, four interference vectors are constructed, 293T cells are co-transfected through the four interference vectors and packaging plasmid, supernatant culture fluid is filtered to obtain virus suspension, chicken embryo fibroblast cell lines and chicken embryo testis sertoli cells are infected through a certain amount of MOI viruses after the virus titer is measured, and the shRNA interference vector with the optimal suppressing efficiency is screened. In the molecular biology, a real-time fluorescence quantification PCR proves that the shRNA interference vector effectively reduces the expression of the chicken cyclin F genes.

Compositions and methods for conferring nematicidal activity to bacteria, plants, plant cells, tissues and seeds are provided. In particular, methods for killing or controlling a nematode pest population, particularly a Pratylenchus spp., e.g., Pratylenchus brachyurus, root knot nematode, reniform nematode, or Lance nematode population, are provided. The methods include contacting the nematode pest with a pesticidally-effective amount of a polypeptide comprising a nematicidal toxin, particularly a nematicidal toxin active against a Pratylenchus spp. nematode, e.g. Pratylenchus brachyurus. Further included are methods for increasing yield in plants by expressing the toxin of the invention.

The methods of the invention relate to methods for preventing testicular infection by bovine viral diarrhea virus by immunizing susceptible male animals against infection.

The invention relates to a method by utilizing rabbit-origin cells to produce swine fever live vaccine, which comprises the following steps that: selecting cells for producing vaccine, preparing cells for producing vaccine, breeding cell virus strain, breeding venom for producing vaccine, preparing vaccine, split charging and freeze drying. The method is characterized in that: rabbit-origin susceptible cells are used for substituting bovine testicular cells or goat kidney cells to produce swine fever live vaccine, which can solve the pollution problem of external virus from cow and sheep, and by strictly controlling the raw materials and the culture conditions, the purity of the produced vaccine can be guaranteed, and the safety and the effect of the vaccine can be guaranteed; and the rabbit-origin susceptible cells to substitute the transferred cells to manufacture the swine fever live vaccine, so the potential hazard that the transferred cells can cause cancer can be avoided, and the safety of the vaccine is higher. The swine fever cell venom cultured through the method has high content of virus, the antigen tilter is 10 times or more of that of the present vaccine, the immune efficacy of the vaccine can be greatly improved, and the weaknesses of the present vaccine in the clinical use can be overcome.

The invention discloses a PCR-RFLP method for distinguishing four types of pratylenchus. A pair of common primers is designed based on the DNA sequence of the D2-D3 region of the 28S gene of nematode ribosome for PCR amplification by DNA extraction, then four types of restriction enzyme are utilized to carry out digestion and band spectrum identification so as to obviously distinguish P. coffeae, P. loosi, P. penetrans and P. vulnus. The PCR-RFLP method has stable results, simple operation, and wide application prospects in the fields of plant quarantine and plant protection.

The invention discloses a biocontrol preparation and application thereof in prevention and treatment of pratylenchus coffea, and belongs to the technical field of microbial pesticides. According to the technical scheme, the biocontrol preparation comprises active ingredients including paenibacillus polymyxa J2-4 and/or a culture and/or metabolite of the paenibacillus polymyxa J2-4; and the paenibacillus polymyxa J2-4 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 21515. The biocontrol preparation is applied to prevention and treatment of the pratylenchus coffea, the problem that in the prior art, a biocontrol preparation which is good in prevention and treatment effect and used for preventing and treating the pratylenchus coffea cannot be provided is solved, and the biocontrol preparation has the advantage that the good prevention and treatment effect on the pratylenchus coffea can be achieved when the biocontrol preparation is used independently.

The invention discloses a kit for preparing transgenic animals through testis injection. The kit for preparing transgenic animals comprises a composition composed of polyethyleneimine and magnetosomes. The kit has the advantages of substantial improvement of the transgenic efficiency, and obtaining of a large amount of transgenic live progenies. The composition in the kit has the advantages of small toxicity, and safety to animals. So the use of the kit in transgenic operation improves the production efficiency of the transgenic animals, and is of great importance in the transgenic animal preparation field.