Subcloning of VHH Single Domain Antibody its transformation into Tobacco mediated Using Agrobacterium (C58GV3101)

摘要

Functional VHH single domain antibody lacking light chains occur naturally in Camelidae. The single domain nature of VHH gives rise to several unique features as compared to antigen-binding derivatives of conventional antibodies. The level of expression in E.coli was found to be too low for therapeutic purposes so we have to use the other production systems like plants. Several Plants are the facile and economic bioreactor for large-scale production of industrial and pharmaceutical agents like proteins and antibodies, and tobacco was our target plant because of large scale production and many other advantages. In this study we have subcloned VHH gene into pBI using phasmid pCANTAB5E. The new construct was used to transform the Agrobacterium strain C58GV3101. Transgenic tobaccos (NC25) Plants were then developed by introducing VHH gene under the control of CaMV 35S promoter. The presence of the VHH antibody gene in the plant genome was verified both by selection of the resistant plants on selective media and by PCR analysis.