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Chinese Journal of Zoonoses ; (12): 203-207, 2015.
Article in Chinese | WPRIM | ID: wpr-474518


SUMOylation is a post‐translational modification involved in various cellular processes .SUMO‐specific protease (SENP) regulates SUMOylation by removing SUMO from conjugated substrates (deSUMOylation) and promoting maturation of SUMO precursor .In order to express Giardia lambia (C2 strain) SENP catalytic domain in E .coli ,the full‐length open reading frame of SENP was amplified by PCR from Giardia lamblia genome DNA .The PCR product about 1 620 bp was cloned into cloning vector pGM‐T .Sequencing result showed the sequence of SENP in C2 strain was identical with that in Gi‐ardia WB strain .Bioinformatics analysis showed that SENP protein possessed a 372 aa discontinuous ULP catalytic domain at C‐terminal .The catalytic domain of SENP was cloned into prokaryotic expression vector pET‐28a(+ ) .The recombinant vector pET‐28a(+ )‐SENPc was transformed into E .coli Rosetta(DE3) ,then the recombinant SENPc protein was expressed by IPTG induction .SDS‐PAGE and Western blot using anti‐His Tag antibody showed that the expression product of SENPc was a fusion protein with a molecular weight of 43 kD .The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SENP protein provide basis for further functional study of SENP .

Article in Chinese | WPRIM | ID: wpr-576084


Objective To provide a basis for further exploration of Dunaliella Salina by determining the main active components in the extract of Dunaliella salina. Methods The carotenoids and vitamins were determined by HPLC or colorimetric method, amino acids were analyzed by amino acid analyze. Unsaturated fatty acid was determined by GC. Atomic absorption spectrophotometry was used to detect K, Na ,Ca, Mg, Cu, Zn, P, Fe, Mn and SPF was used to detect Se. Results There were affluent carotenoids, vitamins, amino acids, unsaturated fatty acid and trace elements in the extract of Dunaliella salina. Conclusion The extract of Dunaliella salina could be used as natural raw material to develop the supplement food.

Article in Chinese | WPRIM | ID: wpr-564087


Aim To investigate whether the polypeptide from Chlamys farreri(PCF)protected HaCaT cells from UVB-induced apoptosis through Fas-caspase-3 and ROS-cytochrome C.Methods Experiment designs were divided into five groups:control group,UVB model group,UVB+5.69 mmol?L-1PCF group,UVB+2.84 mmol?L-1 PCF group,UVB+1.42 mmol?L-1 PCF group.SiRNA for Fas inhibited Fas expression of UVB-induced HaCaT cells.Using agarose gel electrophoresis,the effects of Fas siRNA and ROS scavenger NAC on UVB-induced apoptosis of HaCaT cells were investigated.Expression levels of cytochrome C andcaspase-3 after inhibitory Fas were determined by Western blot analysis.Intracellular ROS was detected by means of an oxidation-sensitive fluorescent probe(DCFH-DA).Results SiRNA for Fas had inhibitory effects on UVB-induced apoptosis of HaCaT cells and caspase-3 expression.NAC had inhibitory effects on UVB-induced apoptosis of HaCaT cells.PCF inhibited UVB-induced generation of ROS and cytochrome C release dose-dependently.Conclusions PCF protected HaCaT cells from UVB-induced apoptosis.Its inhibitory effect on apoptosis could be attributed to inhibition of Fas-caspase-3 and ROS-cytochrome C pathways.

Article in Chinese | WPRIM | ID: wpr-623756


To explore new teaching method of functional experiment,we opened the functional laboratory to encourage students' creativity and train their practical ability and develop their innovative ability.The experiment teaching innovation was attempted and discussed the functional speciality.

Article in Chinese | WPRIM | ID: wpr-591040


Objective To study the effects of polypeptide from Chlamys farreri(PCF) on PI3K/Akt and ASK1-JNK signal transduction pathway in thymocytes after ultraviolet B radiation.Methods Murine thymocytes were exposed to UVB radiation.The reactive oxygen species(ROS) level in thymocytes was detected by colormatching.The activation of Akt was investigated by western-blot.ASK1,JNK activation,mitochondria memberine potential and DNA ladder were also investigated after pretreatment with or without PI3K/Akt pathway inhibitor LY294002.Results PCF could activate Akt,inhibit the activation of the ASK1 apoptosis pathway in murine thymocyte radiated by UVB.Conclusion PCF decreased ROS level,increased Akt activity and inducing ASK1 degradation leading to the inhibition of ASK1-JNK induced apoptosis.