The Expression and Purification of Recombinant Myoglobin and Preparation of Its Quality Control Samples

Abstract

Abstract:

We intended to provide a cheap, stable, highly specific and sensitive lyophilized protein as quality control for diagnositic kits. The myoglobin gene code was optimized and two-step methods were used in this study for the synthesis of myoglobin gene. The DNA fragment of myoglobin was inserted into pET-28a vector forming a recombination plasmid, then, transformed into BL21（DE3）bacteria. Protein expression was induced by 0.5 mmol/L IPTG and purified by Ni Sepharose 6 Fast Flow（FF）. After confirming its specificity, we studied the stability of the lyophilized protein at different storage temperatures（37℃, 25℃, 4℃）. Results showed that the target protein is approximately 20 kD with high specificity, good morphology and stability after freeze-drying under the optimized conditions and protein diluting solution. The lyophilized myoglobin protein with low cost, good stability, high specificity and sensitivity, canbe used as quality control for both qualitative and quantitative myoglobin testing kits.

Key words: Recombinant Myoglobin, Prokaryotic expression, Purification, Lyophilized, Quality control samples

Ren Yanna, Cai Lei, Wu Jianwei, Qian Wei, Zhang Ronghua, Wang Jihua, Tang Shixing. The Expression and Purification of Recombinant Myoglobin and Preparation of Its Quality Control Samples[J]. Biotechnology Bulletin, 2014, 0(8): 189-195.

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