Bitter MAP30 gene expression and cell morphology of BGC823 of.docVIP

 PAGE \* MERGEFORMAT 17 Bitter MAP30 gene expression and cell morphology of BGC823 of Study: Yellow SHAO Shi, Han Xiaohong, Tianshu Wei Han Jun 1, HUANG Shi Teng [Abstract] Objective: To clone the bitter protein MAP30 genes in E. coli BL21 (DE3) in the expression of MAP30 protein in the initial study the impact of BGC 823 cells. Methods: PCR technology from the bitter melon genomic fragment was amplified MAP30 , its identification of TA cloning and restriction enzyme digestion and then sequenced, and recombinant expression vector pET 28a MAP30, transformed into the expression strain BL21 (DE3), the induction by IPTG, with the NTA Ni2 + resin purification of fusion expressed MAP30 protein, and identified by SDS PAGE electrophoresis; purified proteins acting on the BGC 823 cells by light and electron microscopy were observed. Results: The amplified MAP30 gene is 861 bp, and gene bank homology published in DQ643967 100%, and DQ643968 and S79450 homology of 99%; successfully constructed pET 28a MAP30 prokaryotic expression plasmid; by NTA Ni2 + resin purification of the target protein, to obtain the original expression is approximately 30 000 MAP30 fusion protein, and consistent with the forecasts; protein role in cells, the cells have changed significantly. CONCLUSION: The constructed expression vector pET 28a MAP30 and get the original expression of MAP30 fusion protein; recombinant MAP30 protein BGC 823 cells can cause significant morphological changes. [Keywords:] bitter; MAP30; cloning; BGC 823 cells; apoptosis; electron microscope [Abstract] Objective: To clone the MAP30 gene and to express it in E.coli BL21 (DE3) and to detect the influence of the MAP30 protein on proliferation in BGC 823 cells. Methods: Polymerase chain reaction (PCR) was used to amplify the MAP30 gene from bitter guard genome DNA. Then the target gene was inserted into the vector pGEM T. The pGEM T MAP30 vector was transformed into E.coli DH5 @ and identified by restriction dig