Construction Of Plant Express Vector Containing Map30 And Transfer Into Arachis Hypogaea L. And Nicotiana Tabacum L.

MAP30 (a momordica anti-HIV protein of 30kDa) isolated from bitter gourd' s seed was a ribosome-inactivating proteins (RIPs), and was proved anti-HIV, anti-virus, anti-tumor activities and little cytotoxin to normal cells. Until now, there were rarely reports about construction of plant express vector containing map30 and Agrobacterium-mediated map30 transformation. In this experiment, we cloned the map30 gene from bitter gourd, constructed its plant express vector, transferred map30 gene into peanut and tobacco to obtain transgenic plant with anti-virus and anti-microbial activities. in this paper, we added BamHI and Sac I site at the end of map30 gene. The production of PCR cloned into the clonal vector pUCm-T, after sequence analyzed,1 Construction of plant express vector containing map30 gene. Based on the the sequence reported on Genbank, the entire map30 gene including leader sequence was cloned from Momordica charantia (balsam pear) by the use of PCR amplification, and then be subcloned into the binary vector pCAMBIA2303. After the vector pCAMBIA2303-map30 was transformed into Agrobacterium strains GV3101, EHA105, the study on Agrobacterium-mediated transformation of tobacco was carried out. The results from PCR analyse showed that the transformed shoots from the tobacco' s callus confirmed that gene inserted the genome.2 Establishment and optimization of the in planta transformation system of peanut. Confirmed by GUS transient express experiment, a suitable and optimized in planta transformation protocol for peanut would be: after pre-culture for 2d, seeds infected for 7 min with Agrobacterium of OD₆₀₀ 0.3～0.6, then co-culture for 1d～2d. In the transformation experiment of GV3101/pCAMBIA2303-map30, among the 14 cultivar peanut,...