The Analysis Of Real-time PCR For DDK Syndrome Related Gene

ABSTRACT Since the1860s, the DDK inbred strains of mice have shown a "DDK syndrome", a large number of scientists have paid lots of attention to the strange phenomenon of propagation, namely,"as strains within the mating, the DDK males can let inbred females pregnanc normally. However, when DDK females mates with the same subspecies inbred male, the F1fertilized egg deads while blastocyst formation disorders in3-5days of pregnancy, making the DDK female mice almost infertility."Although a large number of scientists based on the phenomenon has been extensively studied, so far the role of the" DDK syndrome "mechanism is not clear.In recent years, the study of the mouse embryo indicates:Expression changes of several categories gene,such as growth faCtors, growth development and apoptosis genes, leukemia inhibitory factor, has a direct and indirect relationship proto-oncogenes with embryonic development,. Preliminary, our laboratory has seleCted eight candidate genes (TGF, C-MYC, LIF, the H-RAS, Mcl-1, Bcl-2and EGF and EGF-R)functional gene as the "DDK syndrome" preliminary exploration, draw:in each period of the DDK (?) x BALB/c (?) mating combinations embryos of Mcl-1, Bcl-2and EGF and EGF-R expression was significantly lowered. So, here we choose the EGF and Bcl-2, Mcl-1, EGF-R four significantly different genes as more in-depth research, and ultimately determine if this gene is related to the DDK syndrome.In this experiment, the mice of clean grade of8-10weeks of age were mated in accordance with the way that male and female ratio is1:1, let (the DDK(?) x BALB/C (?)) as the experimental group mating combination, let (BALB/C(?) the x of BALB /C (?)) mating combination as the control group. Base on the embryonic development time, stage embryos implanted in the early2-cell,4-cell,8-cell were observed and colleCted, and then total RNA was extracted and reverse transcription, and finally make use of quantitative PCR technique, and compare selected candidate genes in different mating combinations embryonic expression differences. The results showed that:EGF and Bcl-2, three of Mcl-1gene expression differences in the three periods are extremely significant; EGF-R gene expression differences in the two periods of the4-cell,8-cell stage is extremely significant, confirmed that the DDK syndrome is related to the role of four candidate genes selected.