- •2) Objects and methods of animal biotechnology
- •3) Totipotent, multipotent, pluripotent animal cells
- •4.Allophenic animals. Genetic chimers
- •5)The principles of genetic cloning
- •6.Allophenic animals. Genetic chimers
- •8) Methods for introducing foreign dna into animal cells
- •9)Cryopreservation of reproductive and germ cells of animals and humans
- •11)The principles and methods of plant cells cultivation in vitro
- •12. The types of medium. Physiological means of compounds medium (as an example you can use the composition of Murashige-Skug medium)
- •14)Differentiation and dedifferentiation in plant cell culture. The obtaining callus mass and cultivation of callus tissue .
- •15)The influence of phytohormons on morphogenesis and regeneration in plant cells culture
- •16.The main path of morphogenesis processes in plant cells culture
- •18.The growth stages in suspension culture
- •20) The factors influenced on microclonal propagation in plant cell culture.
- •21) What is Biotechnology? Various definitions of “Biotechnology”. History of Biotechnology
- •22.Microbial Biotechnology: fundamentals of applied microbiology
- •24.Sterilization in Biotechnology: Methods and principles
- •26) Somaclonal and gametoclonal variation in plant cells culture.
- •27) Artificial seeds". Embryo culture in vitro
- •28. Culture of apical meristem cells
- •29)Cell reconstruction. Theoretical means of cell reconstruction
- •30.Basics of phytopathology. The main diagnostics methods of plant diseases
- •32) Main objects of animal biotechnology:
- •33) Morphological and functional features of gametes - eggs and sperm
- •34Hormonal regulation of mammalian reproduction
- •35)The history of investigations of the genetic transformation of animal cells
- •36.The principles of genetic engineering in animal biotechnology
- •53)Genetic engineering. Methods of genetic transformation
- •54. Methods of receiving plant materials without viruses
- •56) The vector systems used in the genetic engineering
- •57) Methods of genetic engineering: agrobacterial genetic transformation
- •58)Methods of genetic engineering: bioballistics methods
- •60.Apply cell technology and cryopreservation technology for safe gene bank
- •62) Methods of producing chimeras
- •63) Collection and cultivation of oocytes in vivo and in vitro
- •64 Collection and cultivation of embryos in vivo and in vitro
- •66.Fertilization of oocytes in vitro, environment and conditions
- •68) Draw a diagram of the structure of plasmid pBr322
- •69) Draw a diagram of an experiment in genetic engineering (design recDna) and give a description of the main stages
- •70)Describe the calcium-phosphate method for introducing foreign dna into mammalian cells.
- •72 Methods of cryopreservation of sperm and oocytes of mammals
- •74) Modes of freezing and thawing of gametes and embryos
- •75) Methods of artificial fertilization: gamete insemination fallopian tube (gift), zygosity insemination fallopian tubes (zift).
- •76) Stem cells and prospects for their use in practice
- •78.Technical equipment of experiments on artificial insemination
- •80) Methods of animal cloning, reproductive and therapeutic cloning
- •81) Microorganisms in water and wastewater treatment
- •82 Microbial fermentations in food products
- •84.Bacterial examination of water and standard water analysis
- •86) Use of e.Coli for the biotechnological production
- •87) Microbes in milk and dairy products
- •88) What is the benefit of microorganisms in industry
- •90. Algae, their applications
62) Methods of producing chimeras
Chimera is the animal consisting of different genotype cells which originated more than 1 zygotes.
Methods of creating chimeras:
Embryonic aggregation chimeras
Injection of embryonic cells to blastocyst
Therato-carcinomas (for example, the introducton of embryonic cells to the adult)
Xenotransplantation (transplantations of cells, tissue, organs from one adult to another)
63) Collection and cultivation of oocytes in vivo and in vitro
One way to obtain a large number of descendants of genetically valuable livestock females - oocytes fertilization and embryo culture outside the body (in vitro). Traditional methods (in vivo) reproduction of farm animals females produce significantly fewer gametes than males.
The problem of oocyte maturation and fertilization in vitro, followed by culturing embryos and their non-surgical transplant recipients and receiving numerous progeny definitively solved.
Fertilization in vivo matured oocytes in the ovary, when considered in isolation from the maturation of oocytes in vitro, is of limited use for the breeding of farm animals. In this case, as a result of induction of superovulation surgically or during slaughter cows can be obtained in vivo matured units full oocytes. Only a combination of fertilization of oocytes matured in vitro followed by transplantation of embryos can significantly increase the use of the genetic potential of female gametes.
So, if after induction poliovulyatsii per cow - get up to 10 donor tubalnyh, ie matured in vivo, and ovulated oocytes from the ovary is cow can be removed to 200 follicular oocytes. Oocyte maturation by culturing reached 80% or higher, and their fertility - 50-70%.
It is established that in vitro maturation of oocytes not equivalent natural that occurs in vivo in the oocyte prior to ovulation. As shown by numerous studies, the in vitro maturation of the kernel is without the full cytoplasmic maturation. Under the cytoplasmic maturation understand set of morphological, biochemical and physiological changes in the oocyte cytoplasm, resulting in the egg becomes fertilized and capable of further development.
By culturing the oocytes in vitro maturation of immature to understand the process of oocytes in artificial media in which immature oocytes undergo meiotic maturation to metaphase of the second division, that is, to the stage of readiness for fertilization.
To isolate the oocytes from follicles, as a rule, use the ovaries of slaughtered cows and less ovaries extracted operational putem.Posle oocyte retrieval of antral follicle premises in the culture medium is a first morphological changes associated with the nuclear maturation of oocytes, ie meiosis resumes. The first morphological sign of maturity - the destruction of the germinal vesicle - oocyte nuclei, ie prophase oocyte nucleus is converted to a meta-phase.
The first meiotic division, which is a pressure reducer, is replaced by the second equational division in which there is a division of chromosomes in mitosis and spindle formation of ahromatino of new yarns. At this stage of meiotic maturation division is blocked until the oocyte fertilization. On the completion of oocyte maturation in vivo or in vitro judged by the appearance of the polar body or towards the oocyte-mi metaphase of the second division sozrevaniya.Intensivny synthesis, carried out by the oocyte is associated with gene activity and an increase in the number of copies. Found that the first stage of embryonic development take place in an environment where the core does not produce RNA in the cytoplasm, and the process is controlled by RNA accumulated in oogenesis.
The obtained results show that changes in protein synthesis are not associated with nuclear and cytoplasmic maturation of crucial importance for normal fertilization and early embryo development up to the implantation of the embryo in the uterine wall of the recipient. In connection with this, methods for enhancing the synthesis of RNA and protein during maturation of oocytes in vitro. It is assumed that during the maturation of oocytes in vitro in the cytoplasm are not fully synthesized factor responsible for the formation of the male pronucleus.