Homoplasmy stability of transplastomic tobacco plants (Nicotiana tabacum CV. Xhanti) containing human tissue-type plasminogen activator (K2S form) gene in T1 generation

摘要

The rates of recombinant protein in nuclear-transformed plants are often less than 1% of total soluble proteins. As the plant plastid is highly polyploidy, plastid transformation can lead to high-level production of recombinant protein. In addition, plastid transformation has several other advantages such as prevention of gene escape that has a high importance in molecular farming. Tissue-type plasminogen activator (tPA) is an important protein that is used to treating clots in cardiovascular diseases. Thus, production of tPAprotein in plant system was considered. The tPA(K2Sform) gene was transferred to tobacco chloroplast genomes. In this study, we analyzed expression and stability of tPAgene in transplastomic tobacco plants in T1 generation. The presence of tPAgene in transplastomic plants was confirmed with specific primer by PCR analysis. Homoplasmy, gene expression, and protein assay were confirmed with southern blot, RT-PCR, western blotting and ELISA analysis. Results showed that the tPAgene in T1generation of transplastomic tobacco plants is stable and is expressed. In addition, the maximum amount of tPAprotein was estimated up to 0.13% of total soluble protein