Mouse anti-human cTnI monoclonal antibody Fab-gene cloning and sequence analysis of.docVIP

 PAGE \* MERGEFORMAT 7 Mouse anti-human cTnI monoclonal antibody Fab-gene cloning and sequence analysis of Authors: Li Yanyan, Yang Di, Bian Zhi-ping, Xu Jindan, Chen Xiang-jian, Guchun Rong, Zhang Ji-nan [Abstract] Objective: Kelong Shu anti-cTnI mAb Fab fragment gene and sequence analysis. Methods: The design of amplified mouse IgG heavy chain Fd and κ light chain segment primer, from the cTnI mAb secreting hybridoma cell extract total RNA, RT  PCR amplification, molecular cloning of the amplified products, sequencing and sequence analysis. Results: The heavy chain and light chain primers were amplified by an approximately 700 bp and 800 bp DNA fragment. by sequence analysis, and the published mouse IgG gene sequence comparison, the nucleotide and deduced amino acid sequence consistent with mouse IgG1 Fab segmental feature. Log in GenBank, accession number of AY484430 (heavy chain), AY484431 (light chain); amino acid sequence of accession number AAR83243 (heavy chain ), AAR83244 (light chain). Conclusion: The laboratory received a complete mouse-derived anti-human cTnI mAb Fab fragment gene for the mouse anti-human cTnI who laid the foundation for transformation of the source. [Keywords:] antibodies, monoclonal; gene; sequence analysis; serum cardiac troponin [Abstract] AIM: To obtain the gene of murine anti  cTnI Fab fragment and analyse the nucleotide and deduced amino acid sequences. METHODS: By RT  PCR method, the cDNA of anti  cTnI murine antibody Fab fragment was amplified from the hybridoma cells . Cloning and subsequent sequence analysis of the Fab fragment were performed. The deduced amino acid sequence was compared and analysed with previously published sequences. RESULTS: A band of approximate 700 and 800 base pairs was amplified using IgG heavy chain primers and κ light chain primers , respectively. Sequence analysis indicated that the deduced amino acid sequences were in consistent with the characterization of the amino acid prese