Wang Z, Yang D, Wang Q, Li B, Lü Z, Yu J, Zheng H, Fan P, Tang J, Qian M
Shanghai Institute of Biochemistry, Chinese Academy of Sciences, China.
Sci China B. 1995 Sep;38(9):1084-93.
Human interferon-gamma (IFN-gamma) cDNA was synthesized, and it makes the usage of favorable codons in E. coli. The authors got 9 different expression plasmids which contain the synthetic IFN-gamma-cDNA and have different spaces between SD sequence and ATG. The free energies G0f298 in the formation of stable secondary structure in the translation initiation region (TIR) are different in various expression plasmids. One of them, pLY4-gamma 5, can highly yield INF-gamma which will be about 60%-80% of the total bacterial proteins, such a high expression was hardly noted in literature. The reasons of high expression in this work are optimal spaces between SD and ATC, favorable delta G0f298, favourable condons usage for E. coli.
合成了人γ-干扰素(IFN-γ)cDNA,并使其在大肠杆菌中使用偏好密码子。作者获得了9种不同的表达质粒,它们含有合成的IFN-γ cDNA,且SD序列与ATG之间具有不同间隔。不同表达质粒中翻译起始区(TIR)形成稳定二级结构时的自由能G0f298各不相同。其中之一pLY4-γ5能够高效表达IFN-γ,其表达量约占细菌总蛋白的60%-80%,如此高的表达水平在文献中鲜有报道。本研究中高表达的原因在于SD序列与ATG之间的间隔优化、适宜的ΔG0f298以及对大肠杆菌有利的密码子使用情况。