期刊论文

Ma, X.-J.

中文

药学学报

0513-4870

2014

49

1

115-123

10.16438/j.0513-4870.2014.01.006

国家科技支撑计划课题资助项目（2011BAl01803） 国家自然科学基金资助项目（30960500） 广西自然科学基金资助项目（2013GXNSFDA019021,2013GXNSFBA019170）.

本校为其他机构

针对罗汉果果实富含酚类、多糖、油脂和蛋白质等特点, 从3种常用的RNA提取方法中优选出改良Trizol法, 应用该法得到的RNA纯度高(OD_(260/280)= 2.01; OD_(260/230) = 2.02)、完整性好(RIN = 9.50)、得率高(260 μg?g~(-1))。以该法提取的RNA为模板, 通过RACE和RT-PCR技术克隆罗汉果脱氢抗坏血酸还原酶基因全长, 其长度为1 252 bp, 开放阅读框(ORF)为819 bp, 命名为SgDHAR, GenBank登录号为KC907731, 编码一条272个氨基酸的肽链, 理论分子量为30.217 7 kD, 等电点为8.76。该蛋白与GenBank中已登录的DHAR序列比对显示与黄瓜同源性最高(87%)。应用实时荧光定量PCR对SgDHAR在罗汉果不同组织及不同倍...

Total RNA was isolated from Siraitia grosvenorii fruit by the method of modified Trizol, according to S. grosvenorii fruit characteristics of rich phenols, polysaccharide, oil and proteins. The OD260/280, OD260/230, RNA integrity (RIN) and yield of the total RNA with this method were 2.01, 2.02, 9.50 and 260 mirog.g-1, respectively. The open reading frame (ORF) of dehydroascorbate reductase (DHAR), named as SgDHAR, was cloned by rapid amplification of cDNA ends (RACE) and RT-PCR method from S. grosvenorii. The GenBank accession number for this gene is KC907731. The SgDHAR gene contains a full-...