Construction Of Recombined Gene Lat52-DTA And It's Transformation To Arabidopsis And Tobacco

The promoter of a tomato late pollen-specific gene has been constructed with a gene, coding the A chain of Diphtheria toxin, to form a recombined plasmid PLat52-DTA. The plasmid DNA was purified and transformed into Arabidapsis thaliana and tobacco through floral dip and leaf disc transformation to collect transgenic plants serving for the further research work which concentrates on the mutual influences and relationships between vegetative cell and generative cell formed during the process of pollen development. During the binucleate time of pollen development, the mutual influences and relationships between vegetative cell and generative cell have been considered as one of the hot point in understanding the plant generative process. Based on the recombined PLat52-DTA plasmid, two express vectors named as PCB3O2-Lat52-DTA and PBI 101 -Lat52-DTA-GUS were further constructed. After vectors being transformed into tobacco through Agrobacterium-mediated leaf disc transformation method, 38 PPT resistance and 52 Kan resistance transgenic plants were collected after growing on the medium containing antibiotics to be selected for four times. The PCR and Southern analysis are studying now. Further, many untested F2 transgenic seeds of Arabidapsis through floral dip transformation method have been collected, the selection and further molecular analysis is being processed. Moreover, The planting conditions of Arabidapsis have been comfirmed, the diameter of lotus like leaves could grow as long as 8cm or more. Moreover, the transformation methods of it have also been discussed further. The pollen microspore development phages have been identified to the proper floral morphogenesis status through section observations and stains using optical microscope and electron microscope (some data is not shown). These have settled a base for the further cellular observations.