A preliminary study of zinc on the the Drosophila life and polymorphisms in DNA methylation

Zinc known as \DNA methylation Methylation is an epigenetic most in-depth study of a form, is an important mechanism of epigenetic law, widely present in bacteria, plants and higher animals. In higher eukaryotes, DNA methylation regulate important biological functions, including the normal development of the embryo, the maintenance of chromatin structure, X chromosome inactivation, genomic imprinting, gene silencing. Drosophila as one of the most important model organisms in biological research, and its long been considered to be a lack of DNA methylation organisms in recent years, only confirmed the presence of a 5'-methyl cytosine residues in the Drosophila genome DNA methylation levels in early embryonic development the highest overall level of less than invertebrates and plants. Drosophila genomic DNA methylation patterns there are still a lot of controversy, DNA methylation function and mechanism of action remains to be further studied. In addition, many studies reveal that the level of DNA methylation can be influenced by the environment change, and this change is passed to the offspring through epigenetic, thus affecting the DNA methylation pattern of the offspring. Drosophila DNA methylation and environmental factors at home and abroad has not been reported. In this study, Drosophila melanogaster (Drosophila melanogaster) Canton-S strains as experimental material, the improved AFLP markers and statistical analysis was used to detect the different developmental stages and different gender Drosophila genome DNA methylation polymorphism, different concentrations of zinc treatment parental and offspring Drosophila life, fecundity, and at different developmental stages and different gender offspring Drosophila genome DNA methylation polymorphism changes, discusses the different developmental stages and different genders fruit fly genome DNA methylation polymorphism differences, zinc and trace elements of the Drosophila life, fecundity and Drosophila genomic DNA methylation polymorphism, and a preliminary analysis of the effects of zinc progeny in drosophila, fecundity, genetic and genomic DNA methylation polymorphism. The main findings are as follows: 1 instead of MseI endonuclease CPT methylation sensitive AluI as a the AFLP technique of high-frequency shear enzyme, in combination with EcoR I, DNA extraction, double digested connection, pre-amplification a key link in the selective amplification of AFLP technique to optimize the eventual establishment and optimization of AFLP experimental system suitable for the study of Drosophila DNA methylation polymorphism. I.e. EcoR Ⅰ / Alu I restriction enzyme in a total volume of 20μL, the amount of the two enzymes were U, at 37 ° C digestion for 12 h, 65 ° C denaturation 10min; containing 1 U of T4 ligase in 10 ul ligation system, EcoR I joint 5 pmol, Alu I connector 50 pmol 4 ° C connection denaturation at 12 h, 65 ° C 10 min; 25 ul pre-amplification system contains Mg2 1.0 mmol / L, dNTPs 0.2 mmol / L, Taq DNA polymerase 2U, the template 2.5 ul, A-00 50 ng, E-00 50 ng; pre-amplification products were diluted 100-fold selective amplification, the 25 ul selective amplification system containing Mg2 1.0 mmol / L, dNTPs 0.15 mmol / L, Tap DNA enzyme 1.5 U template 2.0μL, E 3 40 ng, A 3 40 ng. From the 64 pairs of selective amplification primers screened with type evenly distributed, with high polymorphism and strong resolution 12 primer pairs to polymorphism of Drosophila DNA methylation. 2.0 3H embryos, 9h embryo, 12 to 15h embryos, 18 to 21h embryos, third instar larvae, pupae at different developmental stages of Drosophila genomic DNA methylation differences of polymorphic bands and more concentrated in the embryonic period embryonic period little difference in polymorphic bands ratio; small number of third instar larvae and pupae DNA methylation polymorphic bands, compared with the embryonic period, the difference was significant; female and male Drosophila genomic DNA amplification the differential bands, indicating that the Drosophila melanogaster DNA methylation there are gender differences, the polymorphic bands ratio was significantly lower than the embryonic period. Medium of appropriate concentrations of zinc (Zn concentration was 2.5%, 5%, 10%) cultured Drosophila, causing the parental (Fo generation of) Drosophila life significantly prolonged, fecundity is also a significant increase compared to the control group; zinc substances concentration is too high (25%) resulted in Drosophila lower life expectancy and loss of reproductive capacity. That the appropriate concentration of zinc material can extend the life of fruit flies, reproductive ability. Medium of appropriate concentrations of zinc (Zn concentration was 2.5%, 5%, 10%) cultured Drosophila, resulting in the F1 generation Drosophila life significantly longer than the control group, a significant increase in fecundity, indicating that the zinc on The Drosophila life and reproductive capacity can be extended to at least the first generation. 5 culture medium to the appropriate concentration of zinc (Zn concentrations were 2.5%, 5%, 10%) cultured Drosophila embryonic offspring 0-3 h, third-instar larvae, pupae and male Drosophila DNA methylation polymorphic strip ratio there was no significant change; 5-8 h embryonic DNA methylation polymorphic bands ratio in the three experimental groups F1-1, F1-2, F1-3 were significantly higher; with the control group than the experimental group, F1-2 (2.5%) of the zinc concentration of zinc material impact on the largest ratio of 5-8 h embryo, 12-15 h embryos, 18-21-h embryos and female flies in DNA methylation polymorphic bands were significantly higher than that in the control group. Progeny Drosophila genomic DNA methylation polymorphisms that the appropriate concentration of zinc material processing can be improved. Drosophila as invertebrate model organisms to study its genetic stability of the external environment factors on life, reproductive capacity, and genomic DNA methylation for functional studies of DNA methylation patterns, methylation and animals Biological Response and protection of environmental stimuli provide important experimental evidence for the human epigenetic and environmental factors and other aspects of the relationship between open new avenues.