ABSTRACT
Objective To construct HSP27 eukaryotic expression plasmids. Methods Full-length HSP27 gene was amplified by reverse transcription polymerase chain reaction (RT-PCR) from breast cancer cell line MCS-7 and cloned into eukaryotic expression vector pAAV-MCS. After the recombinant plasmids transfected into NIH3T3 cells, the expression of HSP27 protein in the host cells was characterized by ECL Western blotting. Results Full-length HSP27 gene was amplified by RT-PCR correctly. The correct cloning of HSP27 gene in pAAV-MCS was confirmed by restriction enzyme digestion and sequencing. ECL Western blotting results indicated that the target gene could express in the mammalian cell line NIH3T3. Conclusion Recombinant plasmid HSP27/pAAV-MCS had been cloned successfully, which would provide the foundation for investigating the role and the mechanism of HSP27 in the ischemia precondition of kidney.
ABSTRACT
Objective To predict the B cell epitope for MUC1 antigen. Methods The secondary structure and surface properties of MUC1, such as physical and chemical characters, hydrophilicity, antigenicity, were analyzed with various methods. Results Many distinct antigenic epitopes in MUC1 were identified by computation: 1-10, 24-54, 65-77, 84-91, 108-134, 140-156, 159-174, 179-196, 199-210, 220-233, 237-265, 270-299, 316-337, 351-362, 369-396, 411-420, and 445-502. Conclusion Prediction of the B cell epitope for MUC1 can provide a base for the studies of structure and function of MUC1, construction of its mutants, and selection of new expression forms of MUC1.