摘要
藤茶叶片富含多酚、多糖以及黄酮类化合物，严重干扰了其总RNA的提取。实验在裂解前先用70％丙酮对藤茶叶片冷冻研磨材料进行多次抽提，然后采用试剂盒提取法、Trizol法和异硫氰酸胍法提取总RNA。结果表明，经丙酮预处理后，上述3种方法均能从藤茶叶片中成功提取到高质量的RNA，其完整性好，纯度高， D260 nm／D280 nm值介于1.8～2.0之间，产率为22.74～33.66μg／g。以提取的总RNA逆转录的cDNA为模板进行RT-PCR，克隆到一条长度为817 bp的pal片段。经Blast比对分析，该基因片段与葡萄、茶树等物种的pal之间具有较高的同源性。
The leaves of Ampelopsis grossedentata are rich of flavones , polyphenols and polymeric carbohydrateds , which hinder the iso lation of RNA.The total RNA with good integrity and high purtity can be extracted from the leaves treated with 70%acetone before ly-sis by the three methods of RNA Extract Kit , Trizol and Guandine thiocyanate .The values of D260 nm/D280 nm of the extracted RNA were between 1.8 to 2.0, the yields were between 22.74-33.66 μg/g FW.A 817 bp phenylalanine ammonia lyase （pal） fragment was successfully amplified from cDNAs reverse-transcribed from the total RNA .By homology analysis ,the cloned pal fragment in Ampelopsis grossedentata is highly identical to the corresponding genes in vitis vinifera and camellia sinensis.
出处
《生物学杂志》
CAS
CSCD
2015年第2期96-99,共4页
Journal of Biology
基金
国家科技支撑计划项目(2013BAD01B05)
福建省自然科学基金(2012J01090)
福建农林大学科技创新平台建设项目(ptjh12015)
关键词
藤茶
叶片
总RNA提取
PAL
Ampelopsis grossedentata
leaf
isolation of total RNA
PAL
