蛋白质内含子在特定氨基酸序列中的剪接  

Inteins Splicing in Specific Amino Acid Sequence

作　　者：张晓[1,2] 刘相钦 孟清[1] 

机构地区：[1]东华大学生物科学与技术研究所,上海201620 [2]加拿大达尔豪斯大学生物化学与分子生物学系,Halifax,Nova Scotia B3H 4R2

出　　处：《中国生物化学与分子生物学报》2016年第6期686-691,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金：加拿大国家科学与工程研究委员会基金;上海市科委攻关项目(No.14521100700);上海市科委国际合作项目(No.14520720200);国家外专局“高端外国专家项目”(No.GDW20143100071);教育部直属高校学校特色项目(No.TS2011DHDX025);国家自然科学基金面上项目(No.31570721;No.31470836);上海市自然科学基金项目(No.13ZR1400600)资助

摘　　要：基因工程技术已经被广泛应用于抗体的生产。但是由于抗体的分子量较大,导致合成抗体较为困难。蛋白质内含子是前体蛋白质中的一段氨基酸序列,能够将自身剪切出来,并将两端的外显子连接形成成熟的蛋白质。将抗体的Fab(antigen-binding fragment)和Fc(crystalline fragment)分别与蛋白质内含子(intein)的N端(I_N)和C端(I_C)融合表达,利用蛋白质内含子的剪接功能,可形成完整的抗体分子。KSCDKTH是存在于抗体铰链区(hinge region)的一段氨基酸序列,如果在KSCDKTH序列中筛选到高效剪接的蛋白质内含子,即可通过蛋白质剪接,将抗体分子的Fab和Fc剪接形成完整抗体。本文筛选发现,Ssp Dna X的3种断裂蛋白质内含子(S0,S1,S11)具有在KSCDKTH序列中高效剪接的能力,这一研究结果为抗体的剪接合成提供了可行性。The genetic engineering has become important ways to produce antibody. Due to its great molecular weight,the antibody is difficult to be produced. Inteins are internal protein sequences capable of catalyzing a protein splicing reaction by self-excising from a precursor protein and simultaneously joining the flanking sequences with a peptide bond. By cleaving the antibody into Fab and Fc fragments and fusing the fragments to the N-terminus and C-terminus of intein respectively,the antibody can be obtained by intein splicing. In order to develop antibody by intein splicing,we must find inteins with high splicing efficiency in KSCDKTH,the amino acid sequence existing in hinge region of antibody. In this article,we found Ssp Dna X-S0/S1/S11 split inteins spliced KSCDKTH sequence successfully,which potentially makes it feasible to develop antibody by inteins splicing.

关 键 词：抗体铰链区 断裂蛋白质内含子 蛋白质剪接 

分 类 号：Q51[生物学—生物化学]