Using The Cpc2 Gene Integration Target Cyanobacteria Transgenic Research

Cyanobacteria show special character in photosynthesis, nitrogen fixation, cytoarchitecture, cytochemistry, metabolization, genetics and evolution. Since this kind of special prokaryote possess very simple genome. the researches of molecular genetic on cyanobacteria have witness significant progress. Particularly in recent years, cyanobacteria have gradually become a very good acceptor system for the expression of foreign gene following the great progress in gene engineering of cyanobacteria.in this experiment, the reconstructed plasmid vector is transformed into Synechococcus sp. PCC7942 by nature transformation. The plasmid pUTK contains cpc homologous fragments cpc2 promoter~. target gene(Ub-Ta1) ~. reporter gene(Kanamycin resistance gene,Kmr)and rbcs-polyA terminator. The foreign gene Ta1 is integrated into chromosome DNA of eyanobacteria by homologous fragments. The kanamycin-resistannt transformants are obtained by kanamycin screening. Southern boning analysis show Ta1 gene have been located at cpc2 gene region. The Ta1 is expressed when cyanbacteria is induced by red light. which confirmed by Western blotting analysis.In the experiments, we have not only analyzed the influence to phycobilisome when foreign gene is integrated into chromosome DNA of Svnechococcus sp. PCC7942 by SDS-PAGE.electron mictoscopic. lectrophoresis. Flow cytometry. but also put forward to the possible mechanism of catching light energy when phycobilisome is destroyed and give a basis for researching into the mechanism of photosynthesis.