广西植物2006，Vol.26Issue(6)：608-616,9.
梅花类黄酮3'-羟化酶基因片段基于基因组DNA的简并PCR法克隆
Cloning of the segment of flavonoid 3′-hydroxylasegene from the gDNA of Prunus mume by degenerate PCR
赵昶灵 1杨清 2陈俊愉1
作者信息
- 1. 南京农业大学,生命科学学院,江苏,南京,210095
- 2. 云南农业大学农学与生物技术学院,云南,昆明,650201
- 折叠
摘要
Abstract
The gDNA was extracted from the tender leaves of P.mume with "PredeimpuritySDS method"developed in this study. Based on the highly conserved amino acid regions of the 11 deduced amino acid sequences of the cDNAs of the flavonoid 3'hydroxylase (F3'H)genes which had been openly published and submitted in the GenBank databases,2 forward degenerate primers and 3 reverse degenerate primers were designed and consisted of 6 sets of primer pair. Only one primer pair could be used in degenerate PCR to amplify three 469 bp segments from the gDNAs of P.mume'Nanjing Hongxu',P.mume'Nanjing Hong'and P.mume'Fenpi Gongfen'respectively. Sequence analysis indicated that the 3 segments shared 99.72% identity among each other and 65.57% identity with the corresponding regions of the 11 cDNAs as a whole. Furthermore,the"GGEK"motif is pointed out not to be the characteristic sequence of F3'H. It is the first time for the segment of F3'H gene to be cloned from the gDNA of ligneous plant. This study can underlay the cloning of the full length of F3'H gene from P.mume.关键词
梅花/基因组DNA提取/类黄酮3'-羟化酶基因片段/简并PCRKey words
P.mume Sieb.et Zucc./extraction of gDNA/segment of flavonoid 3'hydroxylase gene/degenerate PCR分类
农业科技引用本文复制引用
赵昶灵,杨清,陈俊愉..梅花类黄酮3'-羟化酶基因片段基于基因组DNA的简并PCR法克隆[J].广西植物,2006,26(6):608-616,9.
