文献类型：期刊文献

中文题名：角倍总RNA提取方法建立及ACTIN基因片段克隆

英文题名：Extraction of Total RNA from Horned-gall and the Cloning of ACTIN Gene Fragment

作者：阮桢媛[1] 陈晓鸣[1] 杨子祥[1]

第一作者：阮桢媛

机构：[1]中国林业科学研究院资源昆虫研究所,国家林业局资源昆虫培育与利用重点实验室

年份：2012

卷号：25

期号：5

起止页码：551-557

中文期刊名：林业科学研究

外文期刊名：Forest Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2011】；CSCD:【CSCD2011_2012】；

基金：国家林业公益性行业专项(201204602);云南省科技创新强省项目(2010CA028)

语种：中文

中文关键词：角倍;总RNA提取;ACTIN基因

外文关键词：horned-gall; total RNA extraction; ACTIN gene

分类号：Q966

摘要：角倍总RNA的提取是研究角倍蚜虫瘿形成分子机制的基础。本研究针对其多酚、多糖含量高的特点,从各类试剂盒及常规方法改良中优选出CTAB(异丙醇)沉淀方法,较为简单、经济、有效地提取角倍总RNA,其得率可达60.16μg·(100 mg)-1。利用该方法提取的RNA,通过RT-PCR克隆肌动蛋白基因ACTIN片段,碱基序列长度为934 bp,编码311个氨基酸,该序列与GenBank中已登录的ACTIN基因序列比对显示与芒果同源性最高,相似性达到95%,与其它肌动蛋白的氨基酸序列同源性达到86%以上。角倍总RNA的提取及看家基因ACTIN的克隆可为盐肤木和其他蚜虫虫瘿的分子克隆及基因表达分析奠定基础。
The extraction of total RNA from the horned-galls of Rhus chinensis is the basis of the researches on the molecular mechanism of gall formation.Due to high levels of polyphenol and polysaccharides,the modified CTAB method depositing with isopropanol was selected from different kinds of kits and other improved methods.It is a simple,economic and effective method for the extraction of total RNA,and the yield could get 60.16 μg/100 mg.Reverse transcription was conducted on the total RNA extracted by CTAB-isopropanol method,and a 934 bp cDNA sequence of ACTIN gene coding a protein of 311 amino acids was obtained.Homology comparison showed that it shared 95% nucleotide sequence homology with Mangifera indica and over 86% amino acid sequence similarity with actins in other species.The results laid basis for molecular researches of Rhus chinensis and other aphid induced galls.