Polygalacturonase Gene Fragment Cloning Of Processing Tomato And Plant Expression Vector Construction

Processing tomato fruit was used to clone polygalacturonase gene fragment and construct sense and antisense plant expression vectors. The main results were as follows:1. Total RNA was separated from red ripe fruit of processing tomato breeding material BO4 and one fragment about 697bp was cloned by RT-PCR. This fragment was connected with pMD18-T vector by TA cloning to get recombinant plasmid pMD18-T-PG. The fragment was one part of PG gene by sequence analysis.2. An antisense plant expression vector pBinAR-aPG was constructed successfully by inserting the PG gene fragment from pMD18-T-PG digested by Sal I and Sma I into expression vector pBinAR (digested by Sal I and Sma I) between CaMV 35S promoter and OCS terminator.3. The PG gene fragment was cut down from pMD18-T-PG by BamH I and Hinc II and connected with expression vector pBinAR (digested by Sma I and BamH I respectively) and the sense plant expression vector pBinAR-PG was constructed successfully.4. pBinAR-aPG and pBinAR-PG were transferred into Agribacturillus EHA105 respectively by freeze thawing method which could be used for further transformation.