ABSTRACT
We studied the construction of fusion protein TAT-RIG-I-GFP prokaryotic expression vector and verified the function of TAT in transmembrane delivery. First, four pairs of specific primers were designed, and the RIG-I gene of Mallard Duck (Anas platyrhynchos) was cloned. Then, the pET-TAT-RIG-I-GFP and pET-RIG-I-GFP prokaryotic expression vectors were constructed. Meanwhile, they were converted to E. coli BL21 (DE3), which were induced to be expressed after culture. After the purification of His-60 nickel affinity chromatography column and the identification of SDS-PAGE, the purified TAT-RIG-I-GFP and RIG-I-GFP proteins were incubated to DF-1 cells. Finally, fluorescence microscopy was used to observe whether the corresponding fluorescence was produced in DF-1 cells. The results showed that pET-TAT-RIG-I-GFP fusion with TAT showed obvious green fluorescence in DF-1 cells. However, the pET-RIG-I-GFP without TAT cannot display green fluorescence. This shows that TAT-fused protein have successfully delivered DF-1 cells and play a key role in transmembrane delivery. In conclusion, these results provide a solid material basis for further study of antiviral drugs in poultry.
Subject(s)
Cell Membrane , DNA Primers , Escherichia coli , Gene Expression , Gene Products, tat , Genetic Vectors , Recombinant Fusion ProteinsABSTRACT
Objective:To investigate the effects of an atypical antipsychotic drug olanzapine on the body mass index(BMI), leptin and hypothalamic neurohormone related to appetite regulation in the first-attack psychotic patients after 4-week treatment. Methods:Totally 38 first-attack psychotic patients meeting DSM-Ⅳ diagnostic criteria were treated with olanzapine for 4 weeks. The BMI,blood lipids,blood glucose and neurohormone were measured before and after the treatment. The normal control group was also considered the above indices. Results:After the 4-week treatment, the body weight and the body mass index increased significantly when compared with those before the treatment (P < 0.01),and the plasma levels of TC,TG and LDL were higher than those before the treatment(P < 0.05). Both leptin and NPY levels significantly increased in the patients after the treatment (P <0.05),while the NPY levels were significantly lower before the treatment when compared with those in the control group(P< 0.05). The α-MSH levels before and after the treatment were significantly lower than those in the control group(P < 0.01). The CART levels had no significant difference among the pre-treatment group, post-treatment group and control group (P >0.05). Conclusion:The levels of leptin and neurohormone may be changed in the early treatment of olanzapine in the patients with schizophrenia,which may cause weight gain.