The Study Of Construction,identification And Transfection Of Rabbit S100A6 Gene Expression Vector And Its Effect On Neural Cell Proliferation

Objective: Using the method of molecular biology for rabbit Soluble-100A6(S100A6 protein gene)and construct the plasmid expression vector,The target gene expression vector and empty vector were transfected into Schwann Cells(SCs)by liposome mediated transfection,identification of rabbit S100A6 gene expression vector,research S100A6 protein expression in the SCs and the proliferation of SCs.Methods: the sciatic nerve of rabbits was taken,SCs was extracted and cultured.S100A6 gene expression vector plasmid was constructed,relevant verification and identification were carried out for the plasmid,and liposomal mediated transfection technology was used to transfection S100A6 protein c DNA plasmid expression vector and no-loaded plasmid into SCs,namely: group A :S100A6c DNA+SCs.Group B: blank carrier +SCs.After successful transfection,the expression of S100A6 protein in SCs was detected by Western blot,and the changes of S100A6 protein expression in the two groups were analyzed.Meanwhile,immunocytochemistry was used to detect the proliferation of SCs in the two groups.Results: 1.After gel electrophoresis and pires2-egfp-s100A6 double restriction enzyme validation,the S100A6 plasmid was preliminarily identified to be successfully constructed;2.The expression vector plasmid was sequenced by biological company and compared with the standard sequence,confirming the successful construction of S100A6 plasmid vector.2.The expression of S100A6 protein in SCs of the S100A6 overexpression group was significantly higher than that of the control group by Western blot.3.The positive rate of PCNA protein in the overexpression group(38.4% ±4.8%)was significantly higher than that in the control group(8.7%±1.4%),with statistically significant difference(t= 13.44,P<0.001).Conclusion: rabbit S100A6 protein gene expression vector was successfully constructed,and the transfection of SCs could increase the expression of S100A6 protein in cells,and S100A6 protein could promote the proliferation of SCs.