Studied Related To Construct Transgenic Mice Of Green Fluorescent Protein And Visual Of Embryo Engineering

Green fluorescent protein (GFP) from the jelleyfish Aequorea victoria is responsible for the bioluminescence. A Chromophore is formed auto catalytically on the nascent apoprotein's backbone via posttranslational modification. The stable light-stimulated fluorescence is species independent and does not require cofactors, substrates and additional gene products. The signal is detectable by both fluorescence microscopy and eyes. Therefore GFP is a novel and widely applicable genetic marker and protein tag that allow detection in living cells.GFP can elevate preparation of rate in transgenic animal. It will connect both objective gene and GFP gene. Because expression product of GFP gene detect in earlier period of embryonic development, we can only choice and transfer embryo of masculine recombinant of GFP expression. It can reduce aimless of embryo transfer and raise rate of making transgenic animal.GFP gene was introduced into the zygote of FVB/NJ mice by microinjection. The tail of GFP mice obtained from the founders of three weeks mice and picked-up the genome of mice, Genomic DNA was extracted from the offspring of the foundermice for PCR analysis. In conclusion, we confirm the positive transgenic mice. It was attempted to develop green fluorescent protein transgenic mice and produce its lineages. We passage from FO-dynasty transgenic mice to F2-dynasty. We observe birth mice in holo-formatter. There were express of green fluorescent mice. To continue, we gain Fl-dynasty transgenic mice. It was observed to express green fluorescent for 7 mice Fl-dynasty and 35 mice F2-dynasty in holo-formatter. The model of GFP transgenic mice was established primarily. It was important that we could more develop in the visual of embryo engineering of GFP transgenic mice.We carried out three experiments in the visual of embryo engineering of GFP transgenic mice:(l)The oocytes derived from KM strain mice superovulated by conventional method were inseminated with epididymal sperm from transgenic mice of GFP in medium and establish in vitro fertilization model. Then the oocytes were transferred to medium and establish in vitro culture. There were express for green fluorescent after six hours in vitro fertilization.(2) This study was conducted the Embryology of transgenic mice, especially, the activity of embryos in the early embryonic development of the transgenic mice, embryo in medium failed to develop as a result of "2-cell block", but in medium, some of the embryos developed to 4-8-cell stage after culturing two day, and other developed to morula stage. At the same time, we observe to green fluorescent in the morula stage. Embryonic quality is an important factor effecting whether success of assisted reproductive technique, but the decisive factor is the condition of embryo culture in vitro, including component of culture solution and culture system.(3)It was investigated which the embryos of mice were bisected by a fine glass needle attached to a micromanipulator. After zona-softing, the preimplantationembryos of mice were bisected by a fine glass needle attached to a micromanipulator. 2-, 4-, 8-cell embryos and morula were bisected. The half-embryos were cultured in vitro. The success bisecting rate decreased with the increase of cell numbers of embryos, but the rate increased. The half-embryos of 8-cell embryo group and morula group were transfer topseudo pregnant female mice and birth mice.