Single-chain variable fragments with a unique fusion of chemokines lymphoma vaccine prokaryotic expression plasmid and expression of.docVIP

 PAGE \* MERGEFORMAT 26 Single-chain variable fragments with a unique fusion of chemokines lymphoma vaccine prokaryotic expression plasmid and expression of Author: Wangfu Xu Zhao Bing Cheng Yun will be Zhang Xuejun Pan Ling Luo Jian-Min Zuo-Ren Dong Abstract The purpose of this study is to clone mouse B-cell lymphoma cell immunoglobulin (Ig) single-chain variable fragment (scFv), and monocyte chemotactic factor (MCP-3) gene fusion construct fusion scFv-MCP-3 gene expression vector, expression of fusion protein in E. coli, preparation for the treatment of B-cell lymphoma idiotype fusion protein vaccine. Amplified by RT-PCR method using BALB / c mice were the source B-cell lymphoma A20 cell line Ig VH and Ig VL gene, recombinant PCR method using a coding (Gly4Ser) 3 connecting peptide sequence to connect the two genes, preparation of scFv fragments . PCR using the same method of selection of a coding sequence NDAQAPKS connecting peptide linking scFv with the MCP-3 gene, to obtain scFv-MCP-3 fusion gene fragments. Cloned into the prokaryotic expression plasmid pGLo, and expression of fusion protein in E. coli. Sequencing results showed that: A20 cells were successfully cloned Ig VH and Ig VL genes, and successfully prepared scFv fragments and scFv-MCP-3 fusion gene fragment; restriction enzyme digestion method confirmed that the recombinant prokaryotic pGLo/scFv- MCP-3 the fusion gene expression plasmid and accurate insertion. SDSelectrophoresis analysis showed that fusion protein molecular weight of approximately 65 kD, consistent with the expected molecular weight proteins, and the target protein to 30% of total bacterial protein. Conclusion: The successful construction of murine scFv fragments with chemokine MCP-3 fusion B-cell lymphoma idiotype vaccine expression plasmid pGLo/scFv- MCP-3, and to achieve the initial expression of fusion protein. Keywords: purpose of this study is to clone mouse B-cell lymphoma cell immunoglobulin (Ig) sin