The Research Of Human Myoglobin And Aptamer Biosensor

Mb is is one of the most sensitive and specific markers of damaged myocardial. As traditional methods for isolation of Mb from human cardiac tissue are tedious and result in relatively low yield of pure material, so we construct genetic engineering bacteria of recombinant human cardiac Mb with high stability and expression in order to obtain Mb protein as materials use clinical diagnosis and prognosis of my cardiac injury. Tetracycline aptamer biosensor can combine with tetracycline specially and quickly. So it can be used for detecting the concentration of tetracycline in solution.A Pair of Primers was designed and synthesized according to the sequence of Mb gene. The fagment was amplified by PCR from human cardiac total cDNA and inserted into pET-22b(+) to construct an prokaryotic expressing vector. The recombinant was transformed into BL21(DE3) bacteria, and induced by IPTG. The protein was identified by SDS-PAGE showed that Mb gene was expressed successfully after purified by Ni-NTA resin. The expression output of protein was approximately5%of the total bacterial proteins. In the studying of aptamer biosensor, GC was modified with amino. amino GC is covalent with aptamers according to the phosphate group of the oligonucleotide aptamer and amine of GC forming phosphate amine ester covalent bond. Aptamer as recognizable molecular was immobilized on the surface of GC. It can detect tetracycline quickly in samples. Electrical signs from aptamer biosensor can show tetracycline concentration in sample. The lowest bound is1μg/L. The detection time is5minutes.Conclusion, successfully constructing genetic engineering bactria of recombinant Human myoglobin with stability and expression and studying of aptamer biosensor establish a stable base for the early detection of acute miocardial infarction and tetracycline.