摘要
颗粒结合型淀粉合成酶(granule-bound starch synthase, GBSS)是一种能够决定直链淀粉生物合成的关键性酶。根据已发表的多种淀粉粒结合淀粉合成酶(GBSS)基因序列的对比分析,设计一对简并引物,采用同源克隆法和5’/3’RACE (Rapid amplification of c DNA ends)的方法,获得芭蕉芋GBSS全长的cDNA,从芭蕉芋中获得GBSSⅠ基因的cDNA全长片段,并对其核酸序列及其推导的氨基酸序列进行了生物信息学分析,结果表明:芭蕉芋块茎GBSSⅠ基因ORF全长1 851 bp,编码616个氨基酸且蛋白质为一个稳定的蛋白;生物进化分析结果显示芭蕉芋与同姜目下巴西蕉、天宝蕉和马六甲小果野蕉亚种亲缘关系最为接近,芭蕉芋GBSSⅠ的克隆在分子生物学基础上对芭蕉芋块根直链淀粉生物合成及表达途径与调控的研究奠定重要的理论基础。
Granule-bound starch synthase(GBSS) is a key enzyme to determine the amylose synthesis of plants.A pair of degenerate primers were designed according to the conserved region among the known granule_bound starch synthase(GBSS) genes. A full-length of the GBSSI gene c DNA was obtained by Homologous cloning and5’/3’ RACE(rapid amplification of cDNA ends). This study obtain a full-length cDNA fragment of the granule-bound starch synthaseⅠgene from Cannas delis Ker. The nucleotide and deduced amino acid sequences of this gene was analyzed by bioinformatics. The result shows the complete cDNA of GBSSⅠis 1 851 bp, encoding 616 amino acids and the protein is stable;The results of biological evolution analysis show that Cannas delis Ker is most closely related to Brazilian banana, Tianbao banana, and Melaka subspecies banana species. The results indicate that the cloning of GBSSⅠwas of great importance to further studies on the expression and regulation of the key enzymes which were involved in the starch synthesis of leaves from the molecular biology level in Cannas delis Ker.
作者
杨龙
付瑜华
罗春芳
雷静
罗亚红
欧珍贵
Yang Long;Fu Yuhua;Luo Chunfang;Lei Jing;Luo Yahong;Ou Zhengui(Institute of Subropical Crops,Guizhou Academy of Agricultunl Sciences,Xingyi,562400)
出处
《分子植物育种》
CAS
CSCD
北大核心
2020年第22期7376-7384,共9页
Molecular Plant Breeding
基金
贵州省农业科学院基金项目(黔农科院青年基金[2017]09)
贵州省科技支撑项目(黔科合支撑[2018]2291)
农业农村部农业技术试验示范与服务支持项目(滇桂黔石漠化地区特色作物产业发展关键技术集成示范)
贵州省科技计划项目(黔科合平台人才[2019]5804号)
贵州省亚热带作物科技创新人才基地(黔人领发[2016]22号)共同资助
