Preparation of N15 and C13 labeled recombinant protein

With thedevelopment of science and technology, protein structure and metabolism studybegan rapid development, especially in recent years. The NMR (Nuclear Magnetic Resonance)technology has been widely used in the field of protein structure,which becamesimple and feasible to analyze the structure of the protein. The NMR technologyof the basic principle for  analyzing thestructure of the protein is that . the nuclear mass spectrometer issued aseries of electromagnetic wave, stimulating H, N and C atoms, the H, N, C atomsfrom the ground state transition into unstable excited state;when theelectromagnetic wave stopped, excited states of atoms will automatically returnto the ground state, and the excess energy will be released out;by collectingthe energy information and make a further analysis can get the atomic structureof the protein, and draw the protein three-dimensional structure. Although the NMRtechnique is simple, but need protein composed of H, N and C atoms ownning resonanceresonance. The H atom has a natural resonance can be used for analyzing below10kd protein. The conventional protein containing N14 and C12 does not haveresonance, need to adopt their isotope instead. N15 and C13 belongs to thestable isotope which does not own radioactive and are the best substitute forthe onventional protein N14 and C12. So in the process of the protein NMRstructure analysis,the most important aspect is how to obtain a large number ofhigh-quality N15 and C13 labelled protein.

Although there are many ways to get aprotein, accessing to a large number of high-quality protein, still needthrough recombinant protein biosynthesis pathway. On the one hand, easy tocontrol the production conditions, on the other hand the yield and the qualityof the target protein could be guaranteed. Recombinant protein biosynthesis canchoose prokaryotic expression system and yeast expression systems, thebaculovirus insect cell expression system, mammalian cell expression system, butthe target protein need N15 and C13 labelled, which makes the culture ofbacteria and cell culture medium can not be brought into the additional Csource and N source. The recombinant protein was mainly expressed inprokaryotic expression system and yeast expression system, because of itsconvenience, practicality, economy, production, technical difficulty and so on.The prokaryotic expression system commonly used M9 glucose mineral medium toculture of E. Coli;the yeast expression system commonly used czapck medium to culture of yeast .Usually,the C13-labeled monosaccharide or disaccharide are used for the culture mediumC source;the N15-labeled ammonium chloride or sodium nitrate are used for theculture medium N source. The training process of E. Coli and yeast are simpleand the target protein is easy to be purified.

The CLOUD-CLONE CROP has committed to research and develop the production ofrecombinant proteins more than six years, owning complete protein expressionsystems and technology accumulation.We provide a variety of protein synthesis,modification and mark customized services for customer.

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