Screening and identification of cell-penetrating peptides by display technology / 国际药学研究杂志

Screening and identification of cell-penetrating peptides(CPP)which have different functions by display technolo?gy are hot topics in recent years. Currently the most widely used display technology is phage display and mRNA display technology. This paper describes the principle and application of display technology such as phage display and mRNA display and reviews in detail the advance in study on the screening and identification of CPP by display technology in recent years.

Expression and functional analysis of CRT-E2-EGFP fusion protein in B16 cells / 细胞与分子免疫学杂志

AIM: To detect the expression of the Calreticulin and HPV E2 Fusion Protein in B16, and study the effects on proliferation and apoptosis of B16 cell lines in vivo. METHODS: To construct eukaryotic fluoresce expression vector pEGFP-CRT-E2, pEGFP-CRT and pEGFP-E2. Then the recombinant plasmids were transfected into B16 cells by Lipofectamine 2000. The expression of proteins was detected by Western blot. The location of different GFP fusion proteins in B16 was tested by inverted fluoresce microscope. Flow cytometry was applied to detect the effects of fusion proteins on the growing of B16 and then the apoptosis effects of B16 induced by different proteins were observed. RESULTS: The correctly constructed recombinant plasmid pEGFP-CRT-E2 and the expression of CRT-E2 gene could be detected in B16 cells. Apoptosis of B16 cells could be detected after the transient transfection. Meanwhile, the apoptosis rate of B16 cells transfected by pEGFP-CRT-E2 was much higher than that of control cells. And cell cycle G1/G0 arrest was also found (P < 0.01). CONCLUSION: The eukaryotic expression plasmid pEGFP-CRT-E2 is successfully constructed and it could correctly express the fusion protein in B16 cells. And the B16 cells transfected by plasmid pEGFP-CRT-E2 could induce apoptosis.

Vaccination of mice with B16 cells expressing the CRT-E2 fusion protein induces anti-tumor immune responses / 中国免疫学杂志

Objective:To study the anti-tumor effects by vaccination of mice with murine melanoma B16 cells expressing calreticulin (CRT) and Human papillomavirus 16 E2 fusion protein.Methods:Mice were inoculated with B16 cells expressing CRT,E2,or CRT-E2 by intraperitoneal injection. The inoculation was repeated after one week. The activity of CTLs and NK cells,the proliferation of T cells and the amount of IFN-? secreted by spleen lymphocytes were analysed.Results:The activity of CTLs and NK cells,and the amount of IFN-? secreted by spleen lmyphocytes from mice vaccinated with CRT-E2 fusion protein were significantly higher than those of the other groups(P