Abstract
The coumarate:coenzyme A ligase 1 gene (4CL1) was isolated from secondary developing xylem tissues of Pinus massoniana Lamb (Pm4CL1). The Pm4CL1 gene was inserted into a pQE30 expression vector in both sense and antisense orientations to form a pQE30-4CL1-anti4CL1. The expression level of the Pm4CL1 gene in Escherichia coli cells was inhibited by the antisense mRNA of the Pm4CL1 gene. The results of reverse transcriptase polymerase chain reaction and sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the average mRNA and protein expression levels of the Pm4CL1 gene in E. coli cells were decreased by 43 and 46%, respectively. These results are an important basis for further investigation.
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References
Allina SM, Hadash AP, Theilmann DA, Ellis BE, Douglas CJ (1998) 4-Coumarate: coenzyme A ligase in hybrid poplar. Plant Physiol 116:743–754
Antler C (2003) Antisense RNA. Publishing ScqWeb. http://www.scq.ubc.ca/antisense-rna. Issue 4, 2009. Accessed 20 Jul 2009
Atanassova R, Favet N, Martz F, Chabbert B, Tollier MT, Monties B, Fritig B, Legrand M (1995) Altered lignin composition in transgenic tobacco expressing O-methyltransferase sequences in sense and antisense orientation. Plant J 6:465–477
Björn H, Klaus H (2003) The 4-coumarate:CoA ligase gene family in Arabidopsis thaliana comprises one rare, sinapate-activating and three commonly occurring isoenzymes. Proc Natl Acad Sci USA 101:2209–2214
Blee K, Choi JW, Connell AP, Jupe SC, Schuch W, Lewis NG, Bolwell GP (2001) Antisense and sense expression of cDNA coding for CYP73A15, a class II cinnamate 4-hydroxylase, leads to a delayed and reduced production of lignin in tobacco. Phytochemistry 57:1159–1166
Chen ZZ, Chen YC, Chou YH, Lin Y (2006) cDNA cloning and molecular characterization of 4-coumarate:coenzyme A ligase in Eucalyptus camaldulensis. Taiwan J For Sci 21:87–100
Dwivedi UN, Campbell WH, Yu J, Datla RSS, Bugos RC, Chiang VL, Podila GK (1994) Modification of lignin biosynthesis in transgenic Nicotiana through expression of an antisense O-methyltransferase gene from Populus. Plant Mol Biol 26:61–71
Fan BY, Lu H, Jiang XN (2007) High-level expression of 4-coumarate:coenzyme A ligase gene Pt4CL1 of Populus tomentosa in E. coli. For Stud China 9:208–212
Halpin C, Knight ME, Foxon GA, Campbell MM, Boudet AM, Boon JA, Chabbert B, Tollier MT, Schuch W (1994) Manipulation of lignin quality by the down-regulation of cinnamyl alcohol dehydrogenase. Plant J 6:339–350
Hu WJ, Kawaoka A, Tsai CJ, Lung J, Osakabe K, Ebinuma H, Chiang VL (1998) Compartmentalized expression of two structurally and functionally distinct 4-coumarate:CoA ligase genes in aspen (Populus tremuloides). Proc Natl Acad Sci USA 95:5407–5412
Hu WJ, Harding SA, Lung J, Popko JL, Ralph J, Stokke DD, Tsai CJ, Chiang VL (1999) Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees. Nat Biotechnol 17:808–812
Huan HV, Giang HV, Thanh NV, Zhang SL, Wang Y (2010) Identification and functional analysis of the Pm4CL1 gene in transgenic tobacco plant as the basis for regulating lignin biosynthesis in forest trees. Mol Breed. doi:10.1007/s11032-010-9535-9
Kajita S, Katayama Y, Omori S (1996) Alterations in the biosynthesis of lignin in transgenic plants with chimeric genes for 4-coumarate:coenzyme A ligase. Plant Cell Physiol 37:957–965
Lee D, Douglas CJ (1996) Two divergent members of a tobacco 4-coumarate:coenzyme A ligase (4CL) gene family. Plant Physiol 112:193–205
Lee D, Ellard M, Wanner LA, Davis KR, Douglas CJ (1995) The Arabidopsis thaliana 4-coumarate:CoA ligase (4CL) gene: stress and developmentally regulated expression and nucleotide sequence of its cDNA. Plant Mol Biol 28(5):871–884
Lee LK, Roth CM (2003) Antisense technology in molecular and cellular bioengineering. Curr Opin Biotechnol 14:505–511
Pinçon G, Maury S, Hoffmann L, Geoffroy P, Lapierre C, Pollet B, Legrand M (2001) Repression of O-methyltransferase genes in transgenic tobacco affects lignin synthesis and plant growth. Phytochemistry 57:1167–1176
Ratna K, Rashmi V, Manchikatla VR (1998) Potential applications of antisense RNA technology in plants. Curr Sci 74:35–41
Sewalt VJH, Ni W, Blount JW, Jung HG, Masoud SA, Howles PA, Lamb C, Dixon RA (1997) Reduced lignin content and altered lignin composition in transgenic tobacco downregulated in expression of l-phenylalanine ammonia-lyase or cinnamate 4-hydroxylase. Plant Physiol 115:41–50
Shilpakala SR, Savithri HS, Malathi R (2008) Downregulation of gyrase A gene expression in E. coli by antisense ribozymes using RT-PCR. Mol Biol Rep 35:575–578
Voo KS, Whetten RW, O’Malley DM, Sederoff RR (1995) 4-Coumarate:coenzyme A ligase from loblolly pine xylem. Plant Physiol 108:85–97
Wagner A, Donaldson L, Kim H, Phillips L, Flint H, Steward D, Torr K, Koch G, Schmitt U, Ralph J (2009) Suppression of 4-coumarate-CoA ligase in the coniferous gymnosperm Pinus radiate. Plant Physiol 149:370–383
Wei T, Ogbon J, McCoy A (2001) Genetic engineering and lignin biosynthetic regulation in forest tree species. J For Res 12:75–83
Yang XP, Lu H, Chen XM, Jiang XN (2003) Anti-sense 4CL1 changes the content and ratio of lignin and cellulose in transformed tobacco plantlets. J Beijing For Univ 25(3):381–387
Zhang XH, Chiang VL (1997) Molecular cloning of 4-coumarate: coenzyme A ligase in loblolly pine and the roles of this enzyme in the biosynthesis of lignin in compression wood. Plant Physiol 113:65–74
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Huan, H.V., Wang, Y. & Zhang, S. Down-Regulation of Pm4CL1 Gene Expression in E. coli Model by Antisense mRNA. Biochem Genet 50, 529–537 (2012). https://doi.org/10.1007/s10528-012-9497-1
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DOI: https://doi.org/10.1007/s10528-012-9497-1