摘要
为研究漆酶在牛大力生长发育过程中的生物学功能,本实验以牛大力叶片为材料,从反转录PCR获得的cDNA中扩增出漆酶基因CsLAC17全长。序列分析表明,CsLAC17 c DNA全长为2 010 bp,开放阅读框大小为1 755 bp,编码一个由584个氨基酸组成的蛋白质。结构域分析表明,Cs LAC17蛋白的保守结构域具有漆酶典型结构域的特征——铜离子结合域(Cu-oxidase和Cu-oxidase-2)。同源序列分析表明,Cs LAC17蛋白序列与绿豆(Vigna radiata var. radiata)和野生大豆(Glycine soja)同源性为88%、藜豆(Mucuna pruriens)87%、蒺藜苜蓿(Medicago truncatula) 85%。组织特异性表达分析显示,CsLAC17在茎中表达量最高,叶中表达量次之,根中较少。此外,进一步构建了pBI121-CsLAC17过表达载体并转入农杆菌。本研究为日后CsLAC17基因的功能验证以及为牛大力开展分子生物学研究提供帮助。
In this study, the cDNA of LACcase gene Cs LAC17 was amplified by RT-PCR from leaf of Millettia specisoa Champ. Squencing analysis revealed that the full length of Cs LAC17 is 2 010 bp which contains 1 755 bp open reading frame(ORF), and coding a protein consisted by 584 amino acid. Structural domain analysis revealed that the deduced CsLAC17 protein contains copper binding domains(Cu-oxidase and Cu-oxidase-2) which shared the typical characteristics of the LACtase family. Homologous sequence analysis indicated that CsLAC17 protein sequence was 88 percent homologous to Vigna radiata var. radiata and Glycine soja, 87 percent to Mucuna pruriens, and 85 percent to Medicago truncatula. qRT-PCR analysis in different tissues showed that CsLAC17 exhibited the highest expression levels in roots, followed by leaf and less in root. Furthermore, a pBI121-CsLAC17 overexpression vector was constructed, and the plasmid was transformed into Agrobacterium tumefacien. The experiment results will be helpful to the biological function verification of CsLAC17 gene and and to the molecular biology research of C. speciosa.
作者
罗冬梅
赵亚文
荆永琳
徐立
李志英
Luo Dongmei;Zhao Yawen;Jing Yonglin;Xu Li;Li Zhiying(College of Horticulture,Nanjing Agricultural University,Nanjing,210095;Key Laboratory of Crop Gene Resources and Germplasm Enhancementin Southern China,Ministry of Agriculture,Tropical Crops Genetic Resources Institute,Chinese Academy of Tropical Agricultural Sciences,Danzhou,571700;Institute of Tropical Agriculture and Forestry,Hainan University,Haikou,570228)
出处
《分子植物育种》
CAS
CSCD
北大核心
2019年第6期1906-1912,共7页
Molecular Plant Breeding
基金
中央级科研院所基本科研业务费专项(1630032018010)
中央级科研院所非营利启动费专项(pzsfyl-2018-20)共同资助
