Preparation of single chain variable fragment gene in myasthenia gravis

OBJECTIVE?To construct the gene of a single chain variable fragment against the main immunogenic region in acetylcholine receptor.?METHODS?The heavy chain and light chain variable region genes were amplified from an human antigen binding fragment against the main immunogenic region in human acetylcholine receptor using PCR, and cloned in vector phagemid pHEN2. The recombinant phagemid containing the single chain variable fragment gene was subsequently transformed into E coli DH5α. After isolation and purification, the recombinant was digested with endonucleases and checked in agarose gel electrophoresis to confirm the right size of the single chain variable fragment.?RESULTS?The sizes of the heavy chain variable region gene, the light chain variable region gene and the single chain variable fragment gene were found to be approximately 378?bp, 339?bp and 762?bp respectively, and the inserted positions were correct.?CONCLUSION?A single chain variable fragment gene against the main immunogenic region in human acetylcholine receptor has been successfully constructed .