抗骨形成蛋白单链抗体的构建与表达

Construction and expression of bone morphogenetic protein (BMP) single chain Fv

目的：构建抗骨形成蛋白单链抗体并获得表达。方法：应用一段人工合成的含１５个氨基酸的连接肽，采取亚克隆技术，将一株鼠抗ＢＭＰ单克隆抗体的重链可变区（ＶＨ）的Ｎ端和轻链可变区（ＶＬ）的Ｃ端连接起来；将连接产物克隆入融合蛋白表达载体ｐＥＧＸ－４Ｔ－ｌ，在大肠杆菌ＪＭ１０９中进行表达。结果：构建的单链抗体（ＳｃＦｖ）全长７０５ｂｐ，并获得融合表达产物约５２×１０３ｕ。结论：亚克隆方法是一种构建单链抗体快速、简便、可靠的方法。

Objective: To construct and express bone morphogenetic protein (BMP) single chain Fv (ScFv). Methods: A synthetic linder containing 15 amino acids being used, the C end of the heavy chain gene fragment (V 8) and the N end of the light chain gene fragment (V1) of BMP monoclonal antibody were connected with a subclonal method. Then the recombinant BMP ScFv was cloned into pGEX 4T 1 plasmid and induced to express in JM 109 E. coli . Results: BMP ScFv gene consisted of 705 base pairs and the BMP ScFv fusion protein was about 52 kd. Conclusion: Subclonal method is a rapid, simple and reliable way to construct ScFV.