12 results about "Embryonic Stage" patented technology

The invention discloses a comprehensive evaluation method for saline-alkali tolerance of carassius auratus gibelio, which comprises the following steps: performing acute saline-alkali stress on the carassius auratus gibelio and carassius auratus in an embryonic stage, a larva stage and a juvenile stage, performing saline-alkali domestication on a fingerling stage at the same time, increasing salinity and alkalinity once every 10 days in the saline-alkali domestication stage, keeping the up-regulation concentration at a stable maximum value according to fingerling physiological indexes, and obtaining the saline-alkali tolerance of the carassius auratus gibelio. Counting the evaluation value of the saline-alkali tolerance of the carassius auratus gibelio in each stage, determining the weight coefficient of the evaluation value in each stage, then calculating the comprehensive evaluation value of the saline-alkali tolerance in each stage, and comprehensively evaluating the saline-alkali tolerance of the carassius auratus gibelio according to the total value after addition. According to the established saline-alkaline tolerance evaluation system for the carassius auratus gibelio, multiple stages of saline-alkaline tolerance index values are selected for comprehensive weight analysis, the evaluation method is superior to a traditional single-index evaluation method, the tolerance level of the carassius auratus gibelio to saline-alkaline stress can be comprehensively reflected, and basic data are provided for comprehensive evaluation of new germplasm characteristics of the carassius auratus gibelio.

The invention relates to the technical field of aquatic organisms, and particularly discloses a primer pair and method for identifying the genetic sex of Chinese softshell turtles and application of the primer pair. The primer pair comprises 13448-F shown in SEQ ID NO.2 and 13448-Rshown in SEQ ID NO.3. The method comprises the steps that DNA of a sample to be detected is extracted, PCR amplification is conducted through the primer pair, and the sex is judged according to the agarose gel electrophoresis detection result. The primer pair provided by the invention can realize accurate and lossless sex identification in an embryonic stage or a juvenile soft-shelled turtle stage, and provides a key tool for genetic sex control breeding and unisexual fry production of the Chinese soft-shelled turtles.

Disclosed is a sex identification specific DNA sequence and a sex identification method of Chinese mitten crab Eriocheir sinensis. The sex identification specific DNA sequence is showed as SEQ ID NO:1. The sex identification method includes following steps: S1, extracting genomic DNA of Chinese mitten crab Eriocheir sinensis; S2, PCR amplification using the genomic DNA as template; an upstream primer for PCR amplification is as shown in SEQ ID No: 2 and a downstream primer is as shown in SEQ ID No: 3; S3, examining amplified products by agarose electrophoresis, and those with specific amplified bands are females, otherwise they are males. The method has high identification accuracy, is simple and easy to operate, and is suitable for sex identification of Chinese mitten crab Eriocheir sinensis at embryonic and larval stages.

The invention provides application of nicotinamide ribose in preparation of drugs for increasing the number of primordial germ cells and prolonging the growth cycle, and relates to the technical field of cell biology. Animal experiments prove that the supplement of nicotinamide ribose in the embryonic period can significantly increase the number of PGCs and prolong the post-birth growth cycle, and the action mechanism comprises: nicotinamide ribose promotes the proliferation of embryonic cells by up-regulating the NAD + level in PGCs, and enhances the expression of TFAP2C at the same time to maintain the undifferentiated state of cells and delay the meiosis initiation of female embryo PGCs and the differentiation initiation of male embryo PGCs; in post-birth effect evaluation, the number of follicles in ovaries of the female child rats in the NR group is remarkably increased, the fertility cycle is remarkably prolonged, the fertility of filial generations is not affected, and nicotinamide ribose is prompted to effectively maintain reproductive reserve and enhance the fertility by optimizing the development time sequence of PGCs. The invention provides a solid experimental foundation for the application of nicotinamide ribose in increasing the number of primordial germ cells and prolonging the growth cycle, and has a wide application prospect.

The invention discloses a construction method and application of a multi-organ development injury animal model based on non-coding long-chain RNA (Ribonucleic Acid). According to the present invention, it is found that the expression level of LncRNA URS0000A7E830 is significantly increased compared to the normal control group in the embryonic period amoxicillin exposed zebra fish model, and the URS0000A7E830 siRNA microinjection intervention is performed on the fertilized one-cell period zebra fish embryo based on the embryonic period amoxicillin exposed zebra fish model so as to effectively improve the multi-organ differentiation damage. The model established by the invention is novel, reliable, simple and convenient, and has important significance for illuminating the occurrence mechanism of the development injury of multiple organs and determining clinical early warning and intervention targets.

The application discloses a circular RNA circCLASP2 of chicken, a nucleotide sequence of which is shown as SEQ ID NO: 1, and provides detection primers thereof, nucleotide sequences of which are shown as SEQ ID NO: 2-3. The application verifies the existence of a circular transcript circCLASP2 of a CLASP2 gene, proves that the circular transcript is resistant to RNase, and clones the full length of the circular transcript. The application determines through experiments that the circCLASP2 is highly expressed in breast muscle and leg muscle of white-feathered broilers, and is lowly expressed in Dawei mountain chickens; the circCLASP2 is time-sequentially expressed in white-feathered broiler muscle in the embryonic period, and has no obvious change in Dawei mountain chickens. The application verifies that the circCLASP2 mainly exists in cytoplasm through nucleocytoplasmic quantification. The application constructs an overexpression vector of the circCLASP2 and transfects myoblasts, and proves that overexpression of the circCLASP2 can improve the expression of proliferation marker genes and differentiation marker genes, and promote proliferation and differentiation of the myoblasts.

The application discloses a long-chain non-coding RNA molecule lnc46198 related to development of sex glands of semi-muscovy ducks in an embryonic period and application thereof. The sequence of lnc46198 is shown as SEQ ID NO. 1. It is found by the application that lnc46198 is specifically expressed in sex glands of semi-muscovy ducks in an embryonic period, and inhibits proliferation of duck embryo fibroblasts and promotes apoptosis of duck embryo fibroblasts. It is shown that lnc46198 may affect development of sex glands of semi-muscovy ducks in an embryonic period by inhibiting cell proliferation activity, lays a foundation for functions of key lncRNAs in differentiation and development of germ cells of semi-muscovy ducks and the like in the process of differentiation and development of sex glands in an embryonic period, and provides a scientific reference basis for an occurrence mechanism of a sterile trait of semi-muscovy ducks.

The present invention discloses an application of miR-34a-5p and / or target gene NOTCH1 in regulating the growth and development of chicken skeletal muscle, and an application of interfering with miR-34a-5p and / or overexpressing NOTCH1 gene in promoting the growth and development of chicken skeletal muscle. The nucleotide sequence of the miR-34a-5p is shown in SEQ ID NO.1. By verifying the overexpression, interference and targeting relationship of miR-34a-5p, its specific regulatory mechanism in the growth and development of skeletal muscle in the chicken embryonic stage is determined, and the regulatory theory of non-coding RNA in the growth and development of skeletal muscle in the chicken embryonic stage is improved; the NOTCH1 gene is interfered with and functional experiments are performed to determine its specific role in the growth and development of skeletal muscle in the chicken embryonic stage; the miR-34a-5p-NOTCH1 combination disclosed in the present invention can provide a basis for improving the complex regulatory network of chicken skeletal muscle growth and development.

The present disclosure relates to the technical field of prevention and control of diseases in aquatic animals, and particularly relates to a nanocomplex targeting a nervous necrosis virus as well as a preparation method and use thereof. The present disclosure provides a nanocomplex composed of poly(lactic-co-glycolic acid), polyethyleneimine and siRNA. The present disclosure further provides a preparation method of the nanocomplex and use of the nanocomplex in the preparation of a drug for improving the survival rate of fish eggs infected with NNV. In the present disclosure, a vector for an NNV capsid protein is constructed by simulating the NNV; the constructed NNV capsid protein is expressed in cells; the siRNA capable of effectively inhibiting the expression of the viral capsid protein is screened, and then the nanocomplex of the NNV is constructed; and the NNV is blocked in the embryonic stage (fertilized eggs).

The invention provides a precise nutrition regulation and control method for promoting intestinal development of young meat breeding chickens and application, belongs to the technical field of livestock and poultry breeding and production, and aims to solve the problem that hatching eggs laid by meat breeding chickens of 50 weeks old or above are insufficient in nutrition reserve, and L-methionine is injected through amniotic cavities when the meat breeding chickens are hatched to 17 embryonic ages. According to the method, the jejunum length, the jejunum weight and the ileum length (Plt: 0.05) of the 21-day-old offspring broiler chicken are remarkably increased, the expression of ileum proinflammatory factors (IL-8 and TNF-alpha) and apoptosis pathway genes (BAX, Caspase-3 and the like) is down-regulated, the expression of anti-apoptosis genes Bcl-2 is up-regulated, and the serum urea nitrogen level is reduced. Meanwhile, the daily average feed intake and the daily weight gain (Plt; 0.05) are improved. Through accurate intervention in the embryonic period, parent nutrition transfer limitation is broken through, the intestinal development and immune functions of offspring are improved, and an effective solution is provided for healthy breeding of offspring of elderly breeding hens.

The invention relates to the technical field of biology, and particularly discloses a separation and extraction method and application of embryonic-stage mouse spinal microglial cells. The method disclosed by the invention comprises the following steps: taking spinal cord tissues of fetal rats of specific fetal age, and performing steps of fine dissection, non-enzyme temperature dissociation, medium transition, microenvironment regulation and the like to efficiently separate microglia cells which are high in purity and activity and reserve original characteristics of an embryonic period from trace tissues. The method solves the problems of difficult separation, low purity and poor activity of fetal mouse spinal microglia cells in the prior art, and fills the technical blank in the field. The obtained cell can be used for constructing a spinal cord developmental disease model, screening drugs and researching a developmental mechanism, and has important scientific research and clinical application values.

The invention discloses a non-embryonic stage grafting method of monocotyledonous plants and application of grafted bodies of the monocotyledonous plants, and aims to solve the technical problems that the existing monocotyledonous plant embryonic stage grafting technology is extremely short in operation window period, strict in environmental requirement and incapable of being applied in a large scale. The key technical means of the method is that a stock and a scion which are in a non-embryonic development stage (such as a one-leaf one-core stage) are selected, notches are formed in the middle stem parts of the stock and the scion and are in butt joint, and callus formation and vascular connection are promoted through specific fixing and moisturizing treatment. According to the method, the effective grafting time is remarkably prolonged, the dependence on a sterile environment is reduced, the grafting survival rate and the operation feasibility are greatly improved, and an effective means is provided for field large-scale grafting and physiological research of monocotyledonous plants.