title: Expression of the high light-inducible Dunaliella LIP promoter in Chlamydomonas reinhardtii.
abstract: The development of highly inducible promoters is critical for designing effective transformation systems for transgenic analyses. In this study, we investigated the promoter of the light-inducible protein gene (LIP) of the marine alga Dunaliella sp. LIPs are homologs of the early light-induced proteins (ELIPs) of Arabidopsis thaliana. DNA sequence analysis revealed that the LIP promoter contains several light-responsive motifs. Constructs containing progressive truncations of the LIP promoter fused with a Renilla luciferase gene were introduced into Chlamydomonas reinhardtii to identify the light-responsive region in the promoter. Transcription from the LIP promoter was stimulated by high light (HL) in a light intensity-dependent manner. In contrast, oxidative stress induced by chemicals had little effect on the LIP promoter, which implies that the LIP promoter is exclusively induced by high light. Truncation of the promoter to a -100 base pair (bp) region abrogated light inducibility, which suggests the presence of a negative cis-regulatory element upstream of the -100 bp fragment. The LIP promoter can be utilized in transgenic research to specifically select and propagate transgenic microalgae under high-light conditions.
authors.affiliation: Department of Life Science, Hanyang University, Seoul, South Korea.
authors.forename: Seunghye
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Life Science, Hanyang University, Seoul, South Korea.
authors.forename: Yew
authors.initials: Y
authors.lastname: Lee
authors.affiliation: Department of Botany, University of British Columbia, Vancouver, Canada.
authors.forename: Jae-Hyeok
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Department of Life Science, Hanyang University, Seoul, South Korea. esjin@hanyang.ac.kr.
authors.forename: EonSeon
authors.initials: E
authors.lastname: Jin
mesh_terms: D020121:5' Untranslated Regions; D001483:Base Sequence; D016825:Chlamydomonas reinhardtii / Q000235:genetics* / Q000528:radiation effects; D003001:Cloning, Molecular; D018744:DNA, Plant / Q000235:genetics; D017343:Genes, Plant / Q000528:radiation effects; D008027:Light; D010940:Plant Proteins / Q000235:genetics; D011401:Promoter Regions, Genetic / Q000528:radiation effects; D014170:Transformation, Genetic; D016823:Volvocida / Q000235:genetics* / Q000528:radiation effects
chemical_list: D020121:5' Untranslated Regions; D018744:DNA, Plant; D010940:Plant Proteins
keywords: Chlamydomonas; Dunaliella LIP promoter; Heterologous transformation; High light-induced gene expression; Promoter truncation
references.citation: Genetics. 1995 Oct;141(2):543-9
references.pmid: 8647391
references.citation: Proc Natl Acad Sci U S A. 1990 Feb;87(3):1228-32
references.pmid: 2105499
references.citation: Plant J. 2000 Jan;21(2):121-31
references.pmid: 10743653
references.citation: Plant Physiol. 1998 Sep;118(1):227-35
references.pmid: 9733542
references.citation: Proc Natl Acad Sci U S A. 1995 Jan 17;92(2):636-40
references.pmid: 7831342
references.citation: Plant Mol Biol. 2011 Nov;77(4-5):501-11
references.pmid: 21928113
references.citation: J Neurosci. 2002 Jun 1;22(11):4357-63
references.pmid: 12040041
references.citation: Plant Physiol. 2006 Aug;141(4):1264-73
references.pmid: 16778010
references.citation: Photochem Photobiol. 2003 Jun;77(6):668-74
references.pmid: 12870854
references.citation: Int J Mol Sci. 2012;13(3):2676-91
references.pmid: 22489117
references.citation: Genes Cells. 2001 Jul;6(7):607-17
references.pmid: 11473579
references.citation: Mol Cell Biol. 1990 Oct;10(10):5565-8
references.pmid: 2398902
references.citation: Plant Cell. 1997 Apr;9(4):627-40
references.pmid: 9144965
references.citation: Mol Gen Genet. 1999 Oct;262(3):421-5
references.pmid: 10589828
references.citation: Eukaryot Cell. 2008 Jan;7(1):172-6
references.pmid: 17993574
references.citation: Eur J Biochem. 1993 Jun 15;214(3):779-86
references.pmid: 8319687
references.citation: Plant Cell. 1998 May;10(5):673-83
references.pmid: 9596629
references.citation: Proc Natl Acad Sci U S A. 2012 May 15;109(20):E1302-11
references.pmid: 22529359
references.citation: Plant Physiol. 2001 Nov;127(3):986-97
references.pmid: 11706180
references.citation: J Biol Chem. 1991 Jul 25;266(21):13698-705
references.pmid: 1856203
references.citation: Annu Rev Plant Physiol Plant Mol Biol. 1998 Jun;49:525-555
references.pmid: 15012245
references.citation: BMC Genomics. 2008 Nov 26;9:561
references.pmid: 19036138
references.citation: Biochim Biophys Acta. 2001 Nov 1;1506(3):244-59
references.pmid: 11779558
references.citation: Plant Physiol. 1986 May;81(1):92-6
references.pmid: 16664814
references.citation: Mol Cell Biol. 1992 Nov;12(11):5268-79
references.pmid: 1406696
references.citation: Comp Funct Genomics. 2002;3(6):504-10
references.pmid: 18629257
references.citation: EMBO J. 2002 May 15;21(10):2441-50
references.pmid: 12006496
references.citation: New Phytol. 2011 Jun;190(4):896-905
references.pmid: 21299564
references.citation: Plant Physiol. 1995 Mar;107(3):885-893
references.pmid: 12228410
references.citation: Plant Cell Physiol. 2004 Sep;45(9):1221-32
references.pmid: 15509845
references.citation: Plant Mol Biol. 2005 May;58(1):41-51
references.pmid: 16028115
references.citation: Proc Natl Acad Sci U S A. 2010 May 11;107(19):8854-9
references.pmid: 20404166
references.citation: Mol Cell Biol Res Commun. 2000 May;3(5):292-8
references.pmid: 10964753
references.citation: J Exp Bot. 2010;61(1):105-19
references.pmid: 19767307
references.citation: Plant Physiol. 2003 Dec;133(4):1605-16
references.pmid: 14681527
references.citation: Plant J. 2002 Aug;31(4):445-55
references.pmid: 12182703
references.citation: Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3741-6
references.pmid: 10725357
references.citation: Curr Biol. 1995 May 1;5(5):466-8
references.pmid: 7583089
references.citation: Nat Rev Genet. 2007 Mar;8(3):217-30
references.pmid: 17304247
references.citation: Plant Mol Biol. 2000 Jan;42(2):345-51
references.pmid: 10794534
references.citation: Proc Natl Acad Sci U S A. 1988 Oct;85(19):7089-93
references.pmid: 2902624
references.citation: Plant Physiol. 2003 May;132(1):352-64
references.pmid: 12746540
references.citation: Plant Mol Biol. 1998 Nov 1;38(4):633-46
references.pmid: 9747808
references.citation: Plant Cell. 1990 May;2(5):447-55
references.pmid: 2152170
references.citation: Mol Gen Genet. 1995 Oct 25;248(6):727-34
references.pmid: 7476876
references.citation: Mol Genet Genomics. 2001 Jul;265(5):888-94
references.pmid: 11523806
references.citation: Nucleic Acids Res. 1999 Jan 1;27(1):297-300
references.pmid: 9847208
references.citation: Planta. 1998 Jun;205(2):288-96
references.pmid: 9637072
references.citation: Planta. 2001 Sep;213(5):794-801
references.pmid: 11678285
references.citation: Annu Rev Plant Biol. 2009;60:239-60
references.pmid: 19575582
references.citation: Nucleic Acids Res. 2002 Jan 1;30(1):325-7
references.pmid: 11752327
references.citation: Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2610-3
references.pmid: 11607286
references.citation: Plant J. 2000 Oct;24(2):191-203
references.pmid: 11069694
references.citation: Plant Cell. 2003 Aug;15(8):1749-70
references.pmid: 12897250
references.citation: Plant J. 2007 Jun;50(5):795-809
references.pmid: 17553115
references.citation: Plant Cell. 1992 Apr;4(4):485-96
references.pmid: 1498606
references.citation: Mar Biotechnol (NY). 2006 Mar-Apr;8(2):120-8
references.pmid: 16525865
references.citation: Mol Cell Biol. 1989 Sep;9(9):3911-8
references.pmid: 2779571