Familial amyotrophic lateral sclerosis research pGBKT7-mSOD1cDNA yeast expression vector.docVIP

 PAGE \* MERGEFORMAT 11 Familial amyotrophic lateral sclerosis research pGBKT7-mSOD1cDNA yeast expression vector [Abstract] Objective To construct expressed in yeast AH109 mSOD1cDNA shuttle expression plasmid pGBKT7-mSOD1cDNA, of the protein encoded by mutant SOD1 protein - protein interactions. Methods The eukaryotic expression PCR method to set the template pEGFP-mSOD1cDNA, expanding received by the end of 120bp of the mSOD1 gene and the DNA binding domain fragments, after Sal , EcoR restriction enzyme digestion, the gene recombination technique subcloned in the yeast two-hybrid vector pGBKT7 to construct the formation of pGBKT7-mSOD1 eukaryotic expression vector. Results transformation, extraction of plasmid restriction enzyme digestion and nucleotide sequencing, BALST identified that build on the success ratio. Conclusion The construction of shuttle plasmid pGBKT7-mSOD1cDNA, for further study of mutations in the SOD1 gene encoding a protein - protein interactions laid the foundation . [Keywords:] yeast two-hybrid vector subclone mSOD1cDNA pGBKT7 Abstract: Objective To Construct of pGBKT7 mSOD1 Yeast Two Hybrid Expression Vector in study of familial amyotrophic lateral sclerosis and to study Protein protein interactions of mutation SOD1 encode protein. Methods We got the mSOD1 fragment from pEGFP mSOD1 by polymer case reaction (PCR). After digestion by Sal and EcoR , mSOD1 fragment was subcoloned into Yeast Two Hybird expression vector pGBKT7. Results At last, pGBKT7 mSOD1 Yeast Two Hybrid expression vector was constructed and confirmed by Redigestion. Conclusion The construction of pGBKT7 mSOD1 is helpful to the studying of Protein protein interactions . Keywords:: Yeast Two Hybird; expression Vector; subclone; mSOD1CcDNA; pGBKT7 According to SOD (superoxide dismutase, SOD) of different metal, can be divided into three kinds, namely Cu, Zn-SOD, Mn-SOD, Fe-SOD, the body of mammals, only with Cu, Zn-SOD and Mn-SOD. distributed in the cytoplas