摘要
获得功能基因是对植物次生代谢产物代谢途径进行遗传操作的必要条件。通过引入脱氧次黄嘌呤碱基和分解引物,大幅度降低高简并度引物的简并程度,并优化PCR扩增体系和扩增条件,从栀子果实中成功扩增出藏红花素生物合成途径的两个关键酶——玉米黄素剪切加双氧酶(zeaxanthin cleavage dioxygenase,ZCD)和糖基转移酶(glucosyltransferase,GTase)基因的保守区段,分别长170 bp和250 bp。研究结果为今后获得全长基因,并应用于植物藏红花素生物合成途径的调控奠定了基础。
Cloning of key enzyme genes in biosynthesis pathway is significant for modulation of secondary me-tabolite production. Zeaxanthin cleavage dioxygenase （ZCD） and glucosyltransferase （GTase） play important role in crocin biosynthesis pathway in plant. In this paper, we dedicate to clone homologous fragments of ZCDgene and GTase gene from Gardenia fruits. We decreased degeneracy of primers drastically and efficiently through introduction of dI and division of primer groups, and cloned the two enzyme gene fragments successful-ly from Gardenia fruits using optimized RT-PCR. Bioinformatics analysis indicated that the two fragments were 170 bp and 250 bp, encoding conserved regions of ZCD gene and GTase gene, respectively. 3his work will pro-vide a powerful tool for manipulation of crocin biosynthesis pathway in plant in future.
出处
《云南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2012年第5期697-702,715,共7页
Journal of Yunnan Agricultural University:Natural Science
基金
江西省教育厅科学技术研究项目(GJJ11542)
江西中医学院博士启动基金课题(530046)
国家科技支撑计划(2011BAI04B01)
