30 results about "Gonad tissue" patented technology

The invention discloses an Andrias davidianus female specific marker and applications thereof. The steps comprise: A, cloning and verifying a specific fragment: 1) analyzing Andrias davidianus RAD candidate female specific fragments, 2) recovering a PCR product, cloning, and sequencing, and 3) analyzing the cloned and sequenced sequence; B, identifying physiological gender through tissue slicing:1) fixing and storing a collected Andrias davidianus gonad sample, 2) carrying out step-by-step ethanol dehydration on the sample, and 3) slicing the gonad tissue, and observing and identifying the physiological gender of Andrias davidianus; C, establishing a genetic sex identification system: 1) extracting genomic DNA, 2) design specific primers, wherein a nucleotide sequence represented by the verified sequence SEQ ID NO.1 is cloned, and 3) carrying out PCR amplification according to the reaction system; and D, applying the specific molecular marker in the identification of common individuals and sex reversal individuals of Andrias davidianus. According to the present invention, the marker has advantages of simple operation, accuracy and reliability, and can identify the genetic gender of Andrias davidianus through the non-invasive sampling; and the system for identifying the gender of Andrias davidianus and the sex reversal Andrias davidianus is firstly established, the simple and easy-performing molecular biology method is provided for the genetic gender identification of Andrias davidianus, and the approach is provided for the subsequent gender control breeding of Andrias davidianus.

The invention relates to a culture medium for ricefield eel germline stem cells. The culture medium comprises a basic culture medium and FBS, wherein the concentration of the FBS is 5-8%. The invention also relates to a method for separating and culturing the ricefield eel germline stem cells. The method comprises the step of culturing the ricefield eel germline stem cells by using the culture medium. According to the culture medium disclosed by the invention, the adherence efficiency of the ricefield eel germline stem cells can be improved, differentiation of the ricefield eel germline stem cells is prevented, and the proliferation function of the ricefield eel germline stem cells is not influenced. By the method disclosed by the invention, ricefield eel female germline stem cells and male germline stem cells which have higher purity can be separated and obtained in five days by using gonad tissues of ricefield eels. The obtained germline stem cells have typical stem cell morphology,typical stem cell clone can be formed, alkaline phosphatase staining is positive, and related molecular markers of the stem cells are also highly expressed. After the germline stem cells obtained by the method disclosed by the invention are cryopreserved, the germline stem cells can be successfully recovered.

The invention discloses a method for evaluating a PBDEs xenoestrogen effect by using gobiocyprisrarus gene expression, and relates to the evaluation of the influence of water environmental pollution on hydrobios. In the method, the expression quantity of CYP19b genes in the brain tissue and gonad tissue of gobiocyprisrarus cultivated in water with different amounts of PBDEs is detected to illustrate that the PBDEs content of the water is higher, the xenoestrogen effect is more obvious, so that the method for evaluating the PBDEs xenoestrogen effect is obtained.

The invention discloses a method for ascertaining degree of maturity of tridacna ovums. The method comprises the following steps: A, tridacna parent selection: selecting healthy and undamged individual tridacna as an ascertaining object of the degree of maturity of ovums and cleaning and temporarily rearing selected tridacna into seawater; B, collection of gonadal tissues: utilizing an injector tostretch from a tridacna water outlet hole, insert into a gonad surface-layer tissue of tridacna and absorb gonad tissue; C, ascertaining of degree of maturity: performing microscopic observation on the collected gonad tissue. Under a microscope, the observed condition of the degree of maturity of ovums comprises the following three types including an ovum-immaturity stage where ovums are shaped like teardrops or pearls and have the diameters not up to the specification of naturally-discharged ovums; an ovum-maturity stage where ovums are circular or elliptical and have the diameters identicalto ovums in sizes with yolk containing enriched accumulated matter and the ovums have double-layer smooth ovum membranes; and a ovum-maturity stage wherein ovums are circular and the ovum membranes wrinkle so that ovums are likely to crack, thereby making yolk overflow.

The invention relates to the technical field of biology, in particular to establishment and application of a coilia ectenes gonad somatic cell line. The method comprises the following steps: (1) separating and digesting gonad tissues of coilia ectenes, and collecting the cells for plank culturing; and (2) after the cells are adhered to the wall, carrying out passage culture for 45-60 generations in sequence. The obtained coilia ectenes gonad somatic cell line can be stably passaged and proliferated, and can induce fish spermatogonial stem cells to differentiate in vitro to generate sperms.

The invention relates to an early sex identification method for tortoises, belonging to the field of sex differentiation and sex adjustment and control of temperature-dependent sex determination animals. After being weighed, young tortoises, which are two weeks old, are immersed by ethyl alcohol; then, external morphological parameters are acquired and measured; male and female young tortoise gonads are observed; found from tissue section observation, the sexes are determined; external characteristic parameters of the tortoises obviously related to the sexes are obtained through statistical analysis; therefore, a regression equation is obtained; and the sexes of the newborn tortoises can be judged through the regression equation. Animals do not need to be slaughtered; sex identification of the young tortoises, which are about two weeks old, can be carried out through appearance; compared with a gonad tissue section method, the method is simple, high-efficiency and simple to operate; the accuracy rate is higher than 95%; the operability is high; tools required for performing sex identification are simple and easy to obtain; operation is simple; and the tools include a vernier caliper and a computer.

The invention discloses a method for identifying the physiological sex of fish at an early development stage by using 5S rRNA, which comprises the following steps: (1) extracting the total RNA of fish gonad tissues by using a method for conventionally extracting the total RNA of animal cells by using a total RNA extraction kit or a TRIzol reagent, determining an OD260/280 value by using a nucleic acid analyzer, and judging the RNA purity, and then separating total RNA by using an agarose gel electrophoresis method, and observing the brightness and distribution of 5S rRNA, 18S rRNA and 28S rRNA; (2) carrying out microcapillary electrophoresis on the extracted gonad total RNA, and calculating the proportion of 5S rRNA in the total RNA according to the RNA type coverage area obtained by nucleic acid analysis software; (3) according to the proportion of the 5S rRNA in the total RNA, judging the physiological male and female of the fish at the early development stage, when the proportion of the 5S rRNA is smaller than or equal to 9.0%, judging the fish as a testis tissue, and when the proportion of the 5S rRNA is larger than or equal to 12%, judging the fish as an ovarian tissue. The method is easy to implement and simple and convenient to operate, the physiological sex identification of more than 100 fish individuals is realized within 12 hours, and the physiological sex of the fish is determined.

The invention discloses a preparation method of frozen sections of gonad tissues of penaeus vannamei boone. The preparation method comprises the following steps: tissue dehydrating after formalin fixation, tissue embedding, section freezing, section dyeing and sealing. The method provided by the invention is different from a freezing slicing technology of fresh tissues aiming at a freezing slicing technology of penaeus vannamei boone gonad tissues fixed by formaldehyde. Compared with a traditional paraffin slicing method, the method has higher slicing efficiency, and saves the time. Compared with a common freezing slicing method, the method has a better slicing effect and is beneficial to staining observation.

The invention discloses a manufacturing method of a macrobrachium rosenbergii gonad tissue frozen section, a method for manufacturing the macrobrachium rosenbergii gonad tissue frozen section through a frozen section technology and application of the macrobrachium rosenbergii gonad tissue frozen section. The macrobrachium rosenbergii gonad tissue freezing and slicing method mainly comprises the steps of pretreatment of a glass slide before slicing, fixation of tissues by 4% paraformaldehyde, tissue embedding pretreatment, embedding, freezing and slicing and HE dyeing. Compared with a paraffin sectioning technology, the method has higher sectioning efficiency, and compared with a conventional freezing sectioning technology, the method has a more complete tissue form.

The invention discloses a fluorescent protein expression vector, methods for constructing and applying the fluorescent protein expression vector, a sexual gonad tissue specific regulation and expression vector, methods for constructing and applying the sexual gonad tissue specific regulation and expression vector and a method for cultivating ecological safety type transgenic fluorescent ornamental fish. Fluorescent proteins can be driven by the aid of promoters CMV with whole-body general expression functions to be expressed in the ornamental fish, and whole-body expression functions of the ornamental fish can be realized by the aid of the fluorescent protein expression vector. Reverse tetracycline-controlled transactivators rtTA can be driven by sexual gonad specific promoters kop to be expressed, and TRE promoters can be driven to start downstream Cre protein expression plasmid vectors after the reverse tetracycline-controlled transactivators rtTA and tetracycline are combined with one another. Sexual gonad of the ecological safety type transgenic fluorescent ornamental fish cultivated by the aid of the method does not contain transgenic components after the ecological safety type transgenic fluorescent ornamental fish is authenticated.

The invention discloses dsRNA of oncorhynchus mykiss ccne1 gene and application of the dsRNA, and belongs to the technical field of biology. The invention provides a rainbow trout ccne1 gene, an interference gene segment and dsRNA of the rainbow trout ccne1 gene. The method comprises the following steps: firstly, cloning by taking total RNA (Ribonucleic Acid) of gonad tissues of juvenile rainbow trout as a template to obtain a gene sequence of ccne1, wherein the nucleotide sequence of the gene sequence is SEQ ID NO.1, and the amino acid sequence of the gene sequence is SEQ ID NO.2; a ccne1 gene sequence is used as a template to design a primer of an interference gene segment, the interference gene segment is obtained, the nucleotide sequence is SEQ ID NO.3, and the dsRNA of the rainbow trout ccne1 gene is synthesized through in-vitro transcription. The dsRNA of the oncorhynchus mykiss ccne1 gene can efficiently silence the ccne1 gene and effectively reduce the expression of other meiosis related genes in oncorhynchus mykiss, and provides a research idea for oncorhynchus mykiss meiosis pathway analysis.

The invention discloses an identification method for the sex and the gonad maturity degree of apostichopus japonicus, and belongs to the technical field of aquaculture. Parts of gonad tissue of anesthetized apostichopus japonicus are obtained through a negative-pressure tissue collector, the sex and the gonad maturity degree of apostichopus japonicus are identified according to the colors of female and male glands of apostichopus japonicus and the species, the size, the quantity and the structural characteristics of germ cells of female and male glands of apostichopus japonicus. The sex and the gonad maturity degree of apostichopus japonicus are identified with the method, a trauma is small, apostichopus japonicus parents do not need to be dissected, and losses of parents and the economic losses are reduced.

The invention discloses a preparation method of sea urchin peptide oral liquid and aims at solving the problems that sea-urchin gonad can be easily influenced by catching season and the like when being taken as a raw material in the prior art and production cost is high. The preparation method comprises the following steps: selecting sea-urchin gonad tissues, and obtaining RNA of sea-urchin gonad cells by adopting a chloroform extraction method; then adopting an RT-PCR method to obtain cDNA of the sea-urchin gonad cells, then carrying out PCR by utilizing a random primer to obtain all kinds of random gene fragments, then cloning the obtained gene fragments into a prokaryotic expression vector, carrying out induced expression to obtain random protein encoded by a random gene, carrying out protease digestion, and carrying out ultrafiltration to obtain sea urchin peptide solution; and blending the sea urchin peptide solution with wolfberry extract and mint extract to obtain the sea urchin peptide oral liquid. The obtained oral liquid is rich in lycium barbarum polysaccharide, organic acid, glycosides and flavones, taste is mellow, sweetness and acidity are moderate, antioxidant activity is good, and nutritional value is high; and technology is reasonable, operation is easy, production cost is low, and large-scale production can be easily realized.

The present invention discloses a children chest X-ray fixation aid device for imaging departments. The children chest X-ray fixation aid device comprises a bracket, a first radiation shielding plate,connecting rods, a second radiation shielding plate, a sliding mechanism and a fixing mechanism. The children chest X-ray fixation aid device for the imaging departments can conduct a position restraining and fixing treatment on both arms of child patients, in addition, during a chest X-ray process of the child patients, a monitoring end of a chest X-ray apparatus only conducts ray irradiation ofchests of the child patients, heads and gonad tissues of the child parents are not subjected to the ray irradiation by the monitoring end of the chest X-ray apparatus due to shielding of the first radiation shielding plate and the second radiation shielding plate, and thus the device can protect the heads and the gland tissues of the child patients, can also prevent the arms of the child patientsfrom moving to the chest positions and ensures chest X-ray treatment effects of the child patients.

The invention discloses a separation and culture method for germ cells of gonadal tissue of tegillarca granosa. The separation and culture method comprises the following steps: firstly, sucking gonadal tissue of tegillarca granosa by a sterile injector under a sterile condition, carrying out disinfection, performing digestive enzymolysis on the gonadal tissue at 23 DEG C with a mixed solution containing 0.05% of pancreatin and 0.2% of collagenase, collecting a cell suspension, carrying out centrifugation, removing a supernatant, suspending precipitated cells with a disinfection buffer solution, controlling the horizontal centrifugation speed at 2600 rpm/m when taking sperm cells from male gonadal tissue, controlling the horizontal centrifugation speed at 1600 rpm/m when taking egg cells from female gonadal tissue, taking the cell suspension, paving a 6-hole plate, carrying inverted adherent culture in a biochemical culture box at 26 DEG C for 24 h, then setting the plate right, adding1 mL of culture medium to each hole, then, replacing a half of culture medium every 2 d, carrying out constant-temperature standing culture for 5-10 d, and performing primary culture. The separation and culture method selects culture conditions of the gonadal tissue of the tegillarca granosa through a large quantity of basic experiments, so that gonadal cells can be separated, purified, cultured and survive in vitro.

A full-automatic sturgeon gonad tissue puncture sampling method comprises the following steps of preparation before puncture, puncture depth determination through B-mode ultrasound, slitting in an epidermal layer of a specimen for 2-3 mm, puncture position determination, sampling trigger and full-automatic emission gun storage. By adopting the full-automatic sturgeon gonad tissue puncture samplingmethod, sturgeon gonad tissue sampling can be achieved, and the sampling success rate can be also greatly improved.