Cloning and sequencing of the variable region genes of anti-human melanoma monoclonal antibody

摘要

Reverse transcription-PCR (RT PCR) was applied to amplify the variable region genes of the heavy and light chains of a monoclonal antibody(mAb) from a mouse anti-human melanoma hybridoma cell line HB8759. The two genes were sequenced, analyzed by computer and expressed in E. colt. No genes in EMBL Gene Bank 1995were found identical to the VH and VL genes. VH and VL genes consisted of 366 and 324 base pairs respectively.Both VH and VL genes were open reading frames and had antibody characteristics. The two genes were insertedinto expression vector pGEX-4T-1. The results showed that GST-VH and-VL fusion proteins were of anticipated sizes. VH and VL genes were confirmed as functionally rearranged mouse lmmunoglobulin variable region genes.