Study On The Expression Of Pyrroline-5-carboxylate Synthase In Cardiovascular Disease

Objective (s):Study on the purification of Pyrroline-5-carboxylate synthase (P5CS) protein、expression of human P5CS in cardiac tissue、the relationship between P5CS gene expression and disease in dilated cardiomyopathy, coronary heart disease, congenital heart disease.Methods:①The expression of P5CS in human cardiac tissue was studied by Blot Western test, which was collected from the myocardium and atrial septum.②The venous blood of patients with dilated cardiomyopathy, coronary heart disease and congenital heart disease were collected and sequenced by DNA extraction, PCR amplification.③The full-length coding sequence of P5CS(2388bp),was amplified from a human hepatocyte cDNA library by PCR and comfirmed by sequencing. The PCR products were digested with BamH I and Xho I, and ligated into BamH I and Xho I digested pET28a(+)vector. Positive clones was identified. The recombinant plasmid was transformed into E.coli BL21(DE3). were cultured in LB medium and induced with IPTG to protein expression.Results:①P5CS of human express in myocardium and atrial septum.②Currently,gene sequencing did not find the P5CS mutation site from screening of dilated cardiomyopathy,coronary heart disease and congenital heart disease.③P5CS is present in the form of inclusion body, and the target protein is obtained in this study.Conclusion(s):①P5CS express in myocardium and atrial septum.②There was no significant correlation between P5CS and dilated cardiomyopathy,coronary heart disease and congenital heart disease and so on.③P5CS protein was present in the form of inclusion bodies in the expression of Escherichia coli.