作物学报2012，Vol.38Issue(7)：1187-1195,9.DOI:10.3724/SP.J.1006.2012.01187
马铃薯块茎颗粒结合型淀粉合酶基因的克隆及其RNAi载体的构建
Cloning of Granule-Bound Starch Synthase Gene and Construction of Its RNAi Vector in Potato Tuber
摘要
Abstract
There is about 17% starch in potato (Solanum tuberosum L.) tubers. Potato starch granules are composed of two poly-saccharides, 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 gianule-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-VPI-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.关键词
马铃薯/颗粒结合型淀粉合酶/基因克隆/序列分析/RNAi载体Key words
Potato/ GBSSI gene/ Gene cloning/ Sequence analysis/ RNA interference vector引用本文复制引用
刘玉汇,王丽,杨宏羽,余斌,李元铭,张俊莲,王蒂..马铃薯块茎颗粒结合型淀粉合酶基因的克隆及其RNAi载体的构建[J].作物学报,2012,38(7):1187-1195,9.基金项目
本研究由国家科技支撑计划项目(2012BAD06B03),国家现代农业产业技术体系建设项目(CARS-10-P18)和甘肃省重大专项(1102NKDM025)资助. （2012BAD06B03）
