DOI: 10.3724/SP.J.1006.2012.01187

Acta Agronomica Sinica (作物学报) 2012/38:7 PP.1187-1195

Cloning of Granule-Bound Starch Synthase Gene and Construction of Its RNAi Vector in Potato Tuber

There is about 17% starch in potato (Solanum tuberosum L.) tubers. Potato starch granules are composed of two polysaccharides, unbranched amylose (approximately 20% to 25%) and branched amylopectin (approximately 75% to 80%). To develop transgenic potato with high-amylopectin in tubers, we got a cDNA sequence of the potato GBSSI from the total RNA of potato tubers by RT-PCR using specific primers of conserved domain of GenBank Accession Number X58453 sequence. The GBSSI cDNA sequence shared 99.78% similarity with the GBSSI gene in GenBank (accession No. X58453). The full-length of cDNA was 1 824 bp, which contained 607 amino acids, three conserved domains and many important functional sites. The 3D structure of GBSSI was highly similar to that of the glycogen synthase, indicating that GBSSI has a function of starch synthesis. GBSSI similar gene obtained here was granule-bound starch synthase, and its sequence was submitted to GenBank, with the accession number of EU403426. On the basis of a 542 bp RNAi target region from the CDS of GBSSI, the sense and antisense fragments were amplified and separated by a 237 bp intron to construct the RNA interference expression vector pBI121g-PgABI containing “sense gbssA-VP1-ABI3-like protein intron-antisense gbss B” regulated by Patatin promoter, which will lay a solid foundation for the study on synthesis of starch and breeding of transgenic potato with high amylopectin content or pure amylopectin.

Key words:Potato,GBSSI gene,Gene cloning,Sequence analysis,RNA interference vector

ReleaseDate:2014-07-21 16:43:19

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