| The potato (Solarium tuberosum L.) is the fourth food crop and widely uses in the world. It plays very important role in daily life and national economy. Because of its fine processing characteristics, potato starch has extensive range of application, and some application can't be substituted by other source of starch. It is extremely urgent to breed high starch content potato with the rapid development of potato process industry. With he further investigation of relevant enzymes of starch biosynthesis and improvement of molecular biotechnology in recent years, Gene engineering technology for control the activity of relevant enzyme of starch biosynthesis and increasing of starch content of potato became possible.The total RNA extracted from maize (Zea mays L.) endosperm was as template, the coding regions of cDNA of the maize starch branching enzyme gene SBEâ…¡b was successfully amplified by RT-PCR. The result showed that that the cloned fragment has 99% identity to the sequence of the gene in GenBank through BLAST analysis. The coding regions of SBEâ…¡b gene cDNA was 2719bp in length and encode 799 amino acids. According to the reported gene sequence granule-bound starch synthase GBSS gene promoter sequence, the 5'flanking region of granule-bound starch synthase GBSS gene was cloned from the potato cultivar Atlantic. They were a 591bp fragment and an 859bp fragment. One plant over-express vector and the other two plant express-vectors were successfully constructed.Some parameters of Agrobaterium tumefaciens mediated transformation system of potato was gotten by detailed study of Hygromycin B concentration, the pre-incubate time, tincture time and co-culture time. In the experiment, 762 stem sections were infected by mediated of Agrobaterium tumefaciens. Seven transgenic plants were obtained by PCR detected. It proved that the target genes have been integrated into potato genome and can normal express from GUS activity identification of stem sections and potato microtuber. |
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