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Purpose:To clone the vanabie region gene ot light chain ot monoclonal antibody against human retinoblastoma and to analyze the characterization of its nucleotide sequence as well as amino acid sequence.Methods : Total RNA was extracted from 3C6 hybridoma cells secreting specific monoclonal antibody (MeAb) against human retinoblastoma (RB), then transcripted reversely into cDNA with olig-dT primers. The variable region of the light chain (VL) gene fragments was amplified using polymerase chain reaction (PCR) and further cloned into pGEM -T Easy vector. Then, 3C6 VL cDNA was sequenced by Sanger’s method. Homologous analysis was done by NCBI BLAST.Results-. The complete nucleotide sequence of 3C6 VL cDNA consisted of 321 bp encoding 107 amino acid residues, containing four workframe regions (FRs) and three complementarity-determining regions (CDRs) as well as the typical structure of two cys residues. The sequence is most homological to a member of the Vk9 gene family , and its chain utilizes the Jk1 gene
Purpose: To clone the vanabie region gene ot light chain ot monoclonal antibody against human retinoblastoma and to analyze the characterization of its nucleotide sequence as well as amino acid sequence. Methods: Total RNA was extracted from 3C6 hybridoma cells secreting specific monoclonal antibody (MeAb) against human retinoblastoma (RB), then transcripted reversely into cDNA with olig-dT primers. The variable region of the light chain (VL) gene fragments was amplified using polymerase chain reaction (PCR) and further cloned into pGEM-T Easy vector. Then , 3C6 VL cDNA was sequenced by Sanger’s method. Homologous analysis was done by NCBI BLAST. Results -. The complete nucleotide sequence of 3C6 VL cDNA consisted of 321 bp encoding 107 amino acid residues, containing four workframe regions (FRs) and three complementarity- determining regions (CDRs) as well as the typical structure of two cys residues. The sequence is most homological to a member of the Vk9 gene family, and its chain util izes the Jk1 gene