The following figure is a schematic diagram of the culture process of transgenic macaques, please answer according to the figure:
(1) Methods for obtaining green fluorescent protein genes include __________ The core step of genetic engineering is ____
(2) The method of detecting whether the gene of interest is successfully introduced into the recipient cell from the molecular level is ____
(3) The premise of using PCR technology to amplify the gene of interest is to have a known nucleotide sequence of the gene of interest, and to design and synthesize ______
(4) Oocytes collected from female macaques generally need to be cultured until the ________ An important marker for judging egg fertilization is ________
(5) The process (7) in the figure is called ________
【Answer】
Obtaining (1) microinjection of DNA molecule hybridization (paired) primers from the gene library The second division mid -- (MII. mid-stage) In the gap between the egg cell membrane and the zona pellucida can be observed the immune rejection of blastocysts (or mulberry embryos).
【Analysis】
The picture shows the breeding process of transgenic macaques. The role of the marker gene: it is to identify whether the recipient cell contains the gene of interest, so as to screen out the cells containing the gene of interest.
The basic steps of genetic engineering technology: acquisition of the gene of interest→ construction of gene expression vector→ the introduction of the gene of interest into the recipient cell → detection and identification of the gene of interest: detection at the molecular level: (1) detection of whether the DNA of the chromosome of the transgenic organism is inserted into the gene of interest with DNA molecular hybridization technology; (2) to detect whether the gene of interest has transcribed mRNA with molecular hybridization technology; (3) to detect whether the gene of interest is translated into protein with antigen-antibody hybridization technology. Identification at the individual level: insect resistance identification, disease resistance identification, activity identification, etc.
PCR is an abbreviation for polymerase chain reaction, and the principle of amplifying the gene of interest is: the principle of DNA semi-retention replication. Amplification of the gene of interest presupposes a nucleotide sequence of a known gene of interest in order to synthesize (paired) primers based on that sequence. The reaction requires: template DNA, four deoxynucleotides, a pair of primers, and heat-stabilized DNA polymerase (Taqase).
Embryo transfer, also known as fertilized egg transfer, refers to the transfer of early embryos of female animals, or embryos obtained through in vitro fertilization and other means, into the same species of female animals with the same physiological state, so that they can continue to develop into new individuals. Related techniques include: simultaneous estrus, hyperovulation, embryo collection and transfer (in vitro maturation of oocytes, in vitro fertilization of sperm, in vitro fertilization, in vitro culture and transfer of fertilized eggs and a series of other steps).
(1) The green fluorescent protein gene is the target gene, and the methods of obtaining the target gene include: obtaining the target gene from the gene library, amplifying the target gene by USING PCR technology and directly synthesizing it by chemical methods; the core step of genetic engineering is the construction of the gene expression vector, that is, the (1) process in the diagram. (2) The process is to introduce the gene of interest into the recipient cell of the fertilized egg, and the commonly used method is microinjection;
(2) The detection and identification of the gene of interest is divided into detection at the molecular level and identification at the individual level. The method of detecting whether the gene of interest is successfully introduced into the recipient cell at the molecular level is DNA molecular hybridization technology;
(3) The premise of PCR amplification of the gene of interest is that there must be a nucleotide sequence of a known gene of interest in order to synthesize (paired) primers according to this sequence.
(4) The recipient cells often select the denucleated oocytes in the MII( in the second division of the meiolar) stage, and activate them through electrical pulses (or electrical stimulation) to complete the meiotic division and development process; the fertilization process includes 5 stages: 1, sperm through the radiation crown 2, sperm through the zona pellucida 3, sperm into the yolk 4, prokaryosis 5, gamete fusion. The signs of whether fertilization is completed are: in the gap between the yolk membrane and the zona pellucida can be observed two polar bodies;
(5) The process in the figure (7) is embryo transfer, the early embryo used is usually an embryo that develops to the mulberry embryo or blastocyst stage, due to the placental barrier and other reasons, the recipient female monkey will not have an immune rejection reaction to the foreign embryos that move into the uterus, which provides a possibility for the survival of the embryo in the recipient female monkey.