Extraction of high quality of RNA and construction of a suppression subtractive hybridization (SSH) library from chestnut rose (Rosa roxburghii tratt)

被引：21

作者：

Xu, Q

Wen, XP

Tao, NG

Hu, ZY

Yue, HL

Deng, XX ^{[1
]}

机构：

[1] Huazhong Agr Univ, Natl Key Lab Crop Genet Improvement, Wuhan 430070, Peoples R China

[2] Guizhou Univ, Guizhou Key Lab Agr Bioengn, Guiyang, Peoples R China

来源：

BIOTECHNOLOGY LETTERS | 2006年 / 28卷 / 08期

基金：

中国国家自然科学基金;

关键词：

chestnut rose; RNA isolation; Rosa roxburghii Tratt; suppression subtractive hybridization library;

D O I：

10.1007/s10529-006-0020-9

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Chestnut rose (Rosa roxburghii Tratt) is a rare fruit crop of promising economical importance in fruit and ornamental exploitation in China. Isolation of high quality RNA from chestnut rose is difficult due to its high levels of polyphenols, polysaccharides and other compounds, but a modified CTAB extraction procedure without phenol gave satisfactory results. High concentrations of PVP (2%, w/v), CTAB (2%, w/v) and beta-mercaptoethanol (4%, v/v) were used in the extraction buffer to improve RNA quality. The average yield was about 200 mu g RNA g(-1) fresh leaves. The isolated RNA was of sufficient quality for construction of suppression subtraction hybridization (SSH) library, which allowed the isolation of several pathogen-induced defense genes.

引用

页码：587 / 591

页数：5

共 15 条

[1] The presence of CYP79 homologues in glucosinolate-producing plants shows evolutionary conservation of the enzymes in the conversion of amino acid to aldoxime in the biosynthesis of cyanogenic glucosides and glucosinolates [J].