The effectiveness of traditional Chinese medicine (TCM) against various diseases urges more low cost,speed and sensitive analytical methods for investigating the phamacology of TCM and providing a theoretical basis for clinical use.The potential of second-order calibration method was validated for the quantification of two effective ingredients of Schisandra chinensis in human plasma using spectrofluorimetry.The results obtained in the present study demonstrate the advantages of this strategy for multi-target determination in complex matrices.Although the spectra of the analytes are similar and a large number of interferences also exist,second-order calibration method could predict the accurate concentrations together with reasonable resolution of spectral profiles for analytes of interest owing to its ‘second-order advantage'.Moreover,the method presented in this work allows one to simply experimental procedure as well as reduces the use of harmful chemical solvents.

OBJECTIVESTo construct a prokaryotic recombinant vector for mouse lactate dehydrogenase-C and to detect its expression in BL21.

METHODSThe coding sequence of mouse lactate dehydrogenase subunit C was amplified from mouse testis RNA with specific primers, and cloned into pGEX-2T after the restriction digestion with BamH I and EcoR I. GST fusion protein was expressed after induction with IPTG.

RESULTSSequencing and restriction digestion of the recombinant plasmid revealed the existence of coding sequence for mouse lactate dehydrogenase subunit C. A protein band of about 60,000 could be induced by IPTG in the recombinant plasmid.

CONCLUSIONSThe coding sequence of mouse lactate dehydrogenase subunit C was introduced into the pGEX-2T plasmid and a GST-fused protein could be induced at a high level.

Animals ; Escherichia coli ; genetics ; Glutathione Transferase ; genetics ; Isoenzymes ; genetics ; L-Lactate Dehydrogenase ; genetics ; Male ; Mice ; Recombinant Fusion Proteins ; biosynthesis ; Spermatozoa ; enzymology