1.Primary Study On The Effect Of Resveratrol On Early Ovarian Follicle Development And Oocyte Apoptosis In Rat 2.cDNA Cloning And Expression Of Trichomonas Vaginalis RRas Gene

Objective: To study the effect of resveratrol on early ovarian follicle development and oocyteapoptosis in rat.Materials and Methods: The sexual Sprague-Dawley rats were divided into three groups. Theintraperitoneal injection group was the normal neonatal rats which were treated withresveratrol(25mg/kg·d) by intraperitoneal injection. The intragastric administration group wasthe pregnant rats which were treated with resveratrol(25mg/kg·d) by intragastric administrationon day 12 after coition till delivery. Ovaries were collected from the postnatal rats of bothexperimental groups (both intraperitoneal injection group and intragastric administration group)and control group on day 1, 2 and 4 after birth. The ratio of different developmental ovarianfollicles stages was examined by hematoxylin-eosin staining. The oocyte apoptotic profiles weredetermined by terminal deoxynucleotidyltransferase-mediated dUTP nick-end labeling (TUNEL)staining.Results: The ratio of oocytes in oocyte nest decreased and the primordial follicle raised in th1-day-old rats' ovaries of the intragastric administration group. The ratio of oocytes in oocytenest raised and primordial follicle decreased in the 2-day-old rats' ovaries of experimental groups.The ratio of primordial follicles and developing follicles in the 4-day-old rats' ovaries ofexperiment group had no differences with those of control group, while the ratio of oocytesraised. The apoptotic rate of TUNEL in the 2-day-old rats' ovaries of experimental groups had novariance with the one of control group. However, the apoptotic rate of TUNEL in the 1-day-oldrat'ovaries of intragastric administration group was higher than the one of control group.Conclusion: Resveratrol can postpone oocyte nest breakdown and inhibit primordial follicledevelopment, however, it maybe not reduce the apoptosis rate of ovarian follicle. Objective: To clone and express Trichomonas vaginalis RRas gene (TvRRas) for exploring itslocalization and function.Materials and Methods: A cDNA fragment of TvRRas was amplified by PCR, and cloned intopGEM-T easy vector. The recombinant plasmid pGEM-T/TvRRas was digested with restrictionendonuclease BamHI and PstI. The fragment was then subcloned into the expression vectorpET-41a. E.coli BL21 cells were transformed by the recombinant plasmid. The expression wasinduced by IPTG, and the fusion protein was purified with Ni-NTA agarose column and analysedby SDS-PAGE.Result: A cDNA clone with a length of 522 bp was isolated and the recombinant plasmid pET-41a/TvRRas was constructed. The fusion protein (about 45kDa) was expressed and purified.Conclusion: The recombinant plasmid pET-41a/TvRRas have been constructed successfully andthe fusion protein can be expressed in E. coli BL21 cells.