摘要
在Ｅ．ｃｏｌｉ中高表达的中国汉族人源γ干扰素是以包涵体的形式存在的，包涵体经各种溶液洗涤后，用８ｍｏｌ／Ｌ尿素裂解，裂解上清在８ｍｏｌ／Ｌ尿素存在下，经离子交换柱层析和凝胶过滤两步纯化，得到纯度大于９５％的γ干扰素，稀释复性后，其比活为５５×１０５Ｕ／ｍｇ．激光解析电离质谱分析表明，中国汉族人源γ干扰素的分子质量为１７３２ｋｕ，其Ｎ端序列与Ｇｒａｙ等报道的γ干扰素Ｎ端序列一致．中国汉族人源γ干扰素的氨基酸组成分析结果也与其理论值相吻合．
Recombinant human IFN γ from Chinese Han nationality,overexpressed in E.Coli , was found to accumulate in cytoplasmicinclusion bodies.After washing, the inclusion bodies were dissolved in 8 mol/L urea. Under the denatured states, the recombinant human IFN γ from Chinese Han nationality was purified by size exclusion chromatography and ion exchange chromatography.The final yielded product was of high purity(96 87%) and exhibited the mass of molecule 17 32 ku analyzed by mass spectrometry.The renatured IFN γ, which was refolded by diluting, had specific antiviral activity of 5 5×10 5 U/mg. The sequence of 16 amino acid residues from NH 2 terminus of the protein was determined and was found to agree with that of hIFN γ reported by Gray. The composition of amino acids of the protein was analyzed. The result had good agreement with that deduced from cDNA of IFN γ from Chinese Han nationality.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
1998年第6期550-554,共5页
Progress In Biochemistry and Biophysics
基金
国家自然科学基金重点项目
军事医学科学院开发基金
关键词
γ-干扰
中国
汉族人
包涵体
尿素
复性
interferon γ(IFN γ), human IFN γ from Chinese Han nationality, inclusion body, urea, renaturation
