中成药2017，Vol.39Issue(1)：126-130,5.DOI:10.3969/j.issn.1001-1528.2017.01.025
浙贝母肌动蛋白基因的克隆及生物信息学分析
Cloning and bioinformatics analysis of Actin gene in Fritillaria thunbergii
摘要
Abstract
AIM To clone the Actin gene in Fritillaria thunbergii Miq.and to make bioinformatics analysis.METHODS The total mRNA in roots,stems,leaves,flowers and bulbs of F.thunbergii was extracted,and the degenerate primer was designed and synthesized.With total mRNA in leaves as a template,the conserved fragments of Actin gene was cloned by RT-PCR and Ta cloning technology.Using this gene as a reference gene,tissue specificity expression analysis was adopted in 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) gene.RESULTS One gene sequence (463 bp) was obtained by RT-PCR amplification and Ta cloning.The Actin gene in F.thunbergii showed high similarities to those in Lilium regale Wilson,Tulipa gesneriana,Ornithogalum caudatum Jacq.,Dendrobium officinale Kimura et Migo,Diospyros kaki Thunb.,Betula luminifera H.Winkl.and Zea mays L.(84％-98％),the homologies of its amino acid sequence to Drosera adelae F.Muell,Brassica napus L.,Vanilla peanigoeia Ancer,L.regale,Jatropha carcas L.,Lycium barbarum L.and Rhizophora stylosa amino acid sequences were all more than 89％,and the Actin protein had close genetic relationships with Lotus corniculatus L.,L.regale and T.gesneriana.The expressions of HMGR gene in various parts of this plant showed obvious differences,which was in sequence of bulbs ＞ flowers ＞ leaves ＞ stems ＞ roots.CONCLUSION It is the first time that Actin gene (named as FtActin) is coloned in F.thunbergii,which can lay the basis for its effective application.关键词
浙贝母/肌动蛋白(Actin)基因/克隆/生物信息学分析Key words
Fritillaria thunbergii/Actin gene/cloning/bioinformatics analysis分类
医药卫生引用本文复制引用
冯亚斌,施鑫磊,俞信光,沈晓霞,江建铭,王忠华..浙贝母肌动蛋白基因的克隆及生物信息学分析[J].中成药,2017,39(1):126-130,5.基金项目
浙江省中药材育种专项(2016C02058) （2016C02058）
宁波市公益类重大专项(2015C110007) （2015C110007）
浙江万里学院重中之重学科学生创新项目(CX2015002) （CX2015002）
