抗骨形成蛋白(BMP)单链抗体的构建  

CONSTRUCTION OF BMP SINGLE CHAIN ANTIBODY

作　　者：孙远[1] 杨连甲[1] 高玉好[1] 金岩[1] 董绍忠[1] 

机构地区：[1]第四军医大学口腔医学院病理科

出　　处：《免疫学杂志》1997年第3期156-160,共5页Immunological Journal

基　　金：国家自然科学基金

摘　　要：从一株鼠抗ＢＭＰ单克隆抗体杂交瘤中扩增出其可变区基因片段，并将其分别克隆入ｐＵＣ１８、１９载体，利用双脱氧链终止法进行序列测定和计算机分析后，应用一段人工合成的含１５个氨基酸的连接肽，将重链基因（ＶＨ）的Ｃ端和轻链基因（ＶＬ）的Ｎ端连接起来，构建成单链抗体（ｓｃＦｖ）。将其克隆入融合蛋白表达载体ｐＧＥＸ－４Ｔ－１，在大肠杆菌ＪＭ１０９中获得初步表达。Using two sets of oligonucleotide primers which were designed according to FR1 and J region gene sequences of mouse immunoglobulin variable region, we amplified V H and V L gene fragments of BMP McAb by RT PCR. The V H and V L gene fragments were cloned into pUC18,19 vectors, respectively. Being identified by sequencing with Sangers method and analysing with computer, the V H and V L gene fragments were connected by a linker containing 15 amino acids with a subcloning method. Then the recombinant (BMP scFv) was cloned into pGEX 4T 1 plasmid and was expressed in JM 109 E. Coli.

关 键 词：BMP 可变区基因 连接肽 单链 抗体 抗骨形成蛋白 

分 类 号：R392.11[医药卫生—免疫学]