The banana actin 1 promoter drives near-constitutive transgene expression in vegetative tissues of banana (Musa spp.)

被引:14
作者
Hermann S.R. [1 ]
Harding R.M. [1 ]
Dale J.L. [1 ]
机构
[1] Centre for Molecular Biotechnology, School of Life Sciences, Queensland University of Technology, Brisbane, QLD 4001
基金
澳大利亚研究理事会;
关键词
Actin; Banana; Constitutive expression; GUS; Promoter;
D O I
10.1007/s002990100352
中图分类号
学科分类号
摘要
We have isolated a banana actin (ACT1) gene and characterised its expression levels, tissue-specificity and upstream regulatory sequences in banana. The ACT1 promoter (2.2 kbp) was isolated by a combination of ligation-mediated polymerase chain reaction and a method for amplifying flanking sequences. A single transcriptional start site was mapped to within 215 bp of the ATG, within which was located a large intron of 840 bp with almost consensus monocot 5′ and 3′ splice recognition sites. The ACTI gene was amplified from genomic DNA, and the deduced amino acid sequence showed the highest levels of similarity to vegetatively expressed plant actins. Northern analysis indicated that the ACT1 transcript was present in all tissues examined (petals/sepals, stigma/stamens, leaves and roots). Expression studies in transgenic banana plants showed the ACT1 promoter drove strong reporter gene expression in both leaves and roots. Truncations of the ACT1 promoter suggested that all of the necessary regulatory elements required for strong (twofold greater than CaMV 35S) near-constitutive expression were located within 1.2 kbp of the ACT1 ATG.
引用
收藏
页码:525 / 530
页数:5
相关论文
共 34 条
[1]  
Altschul S.F., Madden T.L., Schaffer A.A., Zhang J., Zhang A., Miller W., Lipman D.J., Gapped BLAST and PSI-BLAST: A new generation of protein database search programs, Nucleic Acids Res, 25, pp. 3389-3402, (1997)
[2]  
An Y.-Q., Huang S., McDowell J.M., McKinney E.C., Meagher R.B., The Arabidopsis ACT1/ACT3 actin subclass is expressed in organ primordia and mature pollen, Plant Cell, 8, pp. 15-30, (1996)
[3]  
An Y.-Q., McDowell J.M., Huang S., McKinney E.C., Chambliss S., Meagher R.B., Strong, constitutive expression of the Arabidopsis ACT2/ACT8 actin subclass in vegetative tissues, Plant J, 10, pp. 107-121, (1996)
[4]  
An S.S., Mopps B., Weber K., Bhattacharya D., The origin and evolution of green algal and plant actins, Mol Biol Evol, 16, pp. 275-285, (1999)
[5]  
Becker D.K., Dugdale B., Smith M.K., Harding R.M., Dale J.L., Genetic transformation of Cavendish banana (Musa spp. AAA group) cv 'Grand Nain' via microprojectile bombardment, Plant Cell Rep, 19, pp. 229-234, (2000)
[6]  
Chang S., Puryear J., Cairney J., A simple and efficient method for isolating RNA from pine trees, Plant Mol Biol Rep, 11, pp. 113-116, (1993)
[7]  
Chibbar R., Kartha K., Datla R., Leung N., Caswell K., Mallard C., Steinhauer L., The effect of different promoter-sequences on transient expression of gus reporter gene in cultured barley (Hordeum vulgare L.) cells, Plant Cell Rep, 12, pp. 506-509, (1993)
[8]  
Christensen A.H., Quail P.H., Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants, Transgenic Res, 5, pp. 213-218, (1996)
[9]  
Dugdale B., Beetham P.R., Becker D.K., Harding R.M., Dale J.L., Promoter activity associated with the intergenic regions of banana bunchy top virus DNA-1 to -6 in transgenic tobacco and banana cells, J Gen Virol, 79, pp. 2301-2311, (1998)
[10]  
Dugdale B., Becker D.K., Harding R.M., Dale J.L., Intron-mediated enhancement of the banana bunchy top virus DNA-6 promoter in banana (Musa spp.) embryogenic cells and plants, Plant Cell Rep, 20, pp. 220-226, (2001)