Human Foxm1b prokaryotic expression antibody preparation and its impact on liver cancer cells Foxm1b protein identification.docVIP

 PAGE \* MERGEFORMAT 18 Human Foxm1b prokaryotic expression antibody preparation and its impact on liver cancer cells Foxm1b protein identification Author: Don Paul East, Liu Chun, XU Ya, Zhi-Rong Zeng, Hu Pin-jin 【Abstract】 【Objective】 To construct the human Foxm1b prokaryotic expression vector, expressed and purified protein preparation of polyclonal antibodies, and detection of liver cancer cells using this antibody in Foxm1b gene expression, for further study Foxm1b lay the foundation for gene function. 【Methods】 downloaded from the GenBank genome full-length cDNA sequence of the human Foxm1b and used software to analyze its possible Pcgene epitopes, design primers, applied RT-PCR obtained with epitope Foxm1b gene fragment was cloned into the prokaryotic expression vector pET-28a , in E. coli BL-21 (DE3) was induced to express, application affinity chromatography to obtain high purity protein and prokaryotic expression in rat immune Polyclonal antibody titers detected by ELISA and immunocytochemistry was used to detect This antibody on liver cancer cells of natural Foxm1b protein recognition. 【Results】 Foxm1b successfully constructed recombinant expression vector pET-28a-Foxm1b, purified protein expression in rats after immunization has been a high titer polyclonal antibodies, the antibody can recognize the natural antigen in hepatocellular carcinoma cells. 【Conclusion】 human Foxm1b prokaryotic expression vector, recombinant protein expression, purification and have diagnostic value of antibodies as the basis for subsequent research. Keywords: human Foxm1b gene; prokaryotic expression; polyclonal antibody; hepatoma cell Abstract: 【Objective】 To construct prokaryotic recombinant plasmid and express Foxm1b protein, prepare its polyclonal antibody and apply it to recognize the nature antigen in liver tumor cell line. 【Methods】 The full cDNA sequence of human Foxm1b gene was obtained from GenBank and analyzed by PCgene bioinformatic