Studies On Construction Of MAP30 Plant Expression Vectors And Transformation In Tomato

MAP30 is a 30kDa single-stranded, type-I ribosome inactivating protein (RIP), which possesses anti-tumor and anti-HIV activities. It binds both ribosomal RNA and the HIV-1 long-terminal repeat DNA. In addition to its activity as an N-glycosidase, MAP30 has been reported to (1) inhibit HIV integrase; (2) topologically inactivate viral DNA irreversibly; (3) recognize both DNA and RNA as substrates; and (4) selectively attack tumor transformed and HIV infected cells.MAP30 gene is a special gene without introns, so it can be cloned directly by PCR using balsam pear genome DNA. In this research, two plant expression vectors of MAP30 gene have been constructed, named pBMAP1 and pBMAP2. There is an anchor sequence at the 3'end of MAP30 gene in the pBMAP2, and it can help anchor MAP30 on the Endoplasmic Reticulum in plant cell.The MAP30 gene was introduced into tomato (TX0014) cultivar by Agrobacterium tumefaciens-mediated transformation. The transgenic plants were obtained through kanamycin selection. The presence of MAP30 gene was checked by PCR in six plantlets. Four out of six were confirmed by Southern hybridization. The result of RT-PCR of transgenic plants showed that only three out of four transgenic plants expressed MAP30 mRNA in leaves.As the result, MAP30 gene was treatments and expressed in tomato,and the transgenic plant could be used to produce MAP30 for treatments of anti-HIV and anti-tumor in future.