雨生红球藻β-胡萝卜素酮化酶(bkt)启动子功能分析(英文)
FUNCTIONAL ANALYSIS OF THE PROMOTER OF BKT ENCODING BETA-CAROTENE KETOLASE IN HAEMATOCOCCUS PLUVIALIS
-
摘要: 单细胞绿藻---雨生红球藻在逆境条件下积累大量的虾青素。β-胡萝卜素酮化酶(bkt)催化在β-胡萝卜素和玉米黄素的β-紫罗酮环C-4位引入酮基的反应,是虾青素合成过程中的关键酶。我们利用凝胶阻滞的方法研究雨生红球藻中bkt基因309bp(-617/-309)启动子区域的转录因子结合位点并发现在-396/-338的59bp探针存在特异的核蛋白结合位点。通过序列分析,发现此59bp区域并不包含TATA或者CAAT-box,而是存在对光、缺氧、p-香豆酸及激素反应的G-box。Abstract: The unicellular green alga Haematococcus pluvialis accumulates a highvaluable astaxanthin under stress conditions. Betacarotene ketolase (BKT), a key enzyme in astaxanthin biosynthesis in H. p luvialis, catalyzes the conversion of B-carotene to canthaxanthin and zeaxanthin to astaxanthin. Electrophoresis mobility shift assay (EMSA) was used in H. pluvialis to identify transcription factor binding sites within a 309 bp promoter region (-617/-309) of betacarotene ketolase gene and a 59 bp sequence between -396 and -338 bp was found to have a specific binding activity to the nuclear protein. Sequence analysis revealed that this important functional region contains neither TATA nor CAAT box but a Gbox involved in the responsiveness of light, anaerobiosis, pcoumaric acid and hormone.
-
Keywords:
- EMSA /
- Haematococcus pluvialis /
- bkt /
- Promoter /
- Transcription factor binding sites
-
-
[1] Manjunath S, Sachs M M. Molecular characterizat ion and promot eranalysis of the maize cytosol ic glyceraldehyde 3-phosphate dehydrogenasegene family and its expression during anoxia [J]. Plant Mol ecularBiology, 1997, 33(1) : 97 ) 112
[2] Lotan T, Hirschberg J. Cloning and expression in Escherichia col i ofthe gene encoding beta-C-4-oxygenase, that converts beta-carot ene tothe ketocarotenoid canthaxanthin in Haematococcus pluvialis [J].FEBS letters, 1995, 364: 125
[3] Grnewald K., Hirschberg J., Hagen C. Ketocarotenoid biosynthesisoutside of plastids in the unicellular green alga Haematococcus pluvialis[J]. The Journal of Biological Chemistry, 2001, 276: 6023 ) 6029
[4] Kajiwara S, Kakizono T, Sait o T, et al. Isolation and functionalidentificat ion of a novel cDNA for astaxant in biosynthesis fromHaematococcus pluvialis, and astaxanthin synthesis in Escherichia coli[J]. Plant Molecular Bi ology, 1995, 29: 343 ) 352
[5] Linden H. Carotenoid hydroxylase from Haematococcus pluvialis :cDNA sequen ce, regulat ion and funct ional complement ation [J].Biochimica et Biophysica Acta, 1999, 1446: 203) 212
[6] St einbrenner J, Linden H. Regulation of two carotenoid biosynthesisgenes coding for phyt oene synthase and carotenoid hydroxylase duringstress-induce astaxanthin format ion in the alga Haematococcus pluvialis[J]. Plant Physiology, 2001, 125: 810) 817
[7] Sun Z, Francis X, Cunningham J, Gantt E. Diff erential expressionof two isopent enyl pyrophosphate isomerases and enhanced carotenoidaccumulation in a uni cellular chlorophyt e [J]. Proceedings of NationalAcademy of Sciences USA, 1998, 95: 11482 ) 11488
[8] Grnewald K, Eckert M, Hirschberg J, et al.Phytoene desaturase islocalized exclusively in the chloroplast and up-regulat ed at the mRNAlevel during accumulat ion of secondary carot enoids in Haematococcuspluvialis (Volvlcales, chlorophyceae) [J]. Plant Physi ology, 2000,122: 1261) 1268
[9] Meng C X, Teng C Y, Jiang P, et al. Cloning and characterizat ionof beta-carotene ketolase gene promoter in Haematococcus pluvialis7 50 ACTA HYDROBIOLOGICA SINICA 30卷[J]. Acta Biochimica Bi ophysica Sin, 2005, 37(4) : 270) 275
[10] Lescot M, Dehais P, Thi js G, et al. PlantCARE: a database ofplant cis-act ing regulatory elements and a portal to tools for in sil icoanalysis of promoter sequences [J]. Nucleic Acids Research, 2002,30: 325 ) 327
[11] Rydz S K, Prieto J L, Rychter A M, et al. A DNA-binding activityfor the promoter of the gene encoding C4 phosphoenolpyruvate carboxylaseis modulat ed by phosphorylation during greening of theSorghum leaf [J]. Plant Science, 2000, 159: 65 ) 73
[12] Giuliano G, Pichersky E, Malik V S, et al. An evolutionarily conservedprotein binding sequence upstream of a plant light regulatedgene [J]. Proceedings of National Academy of Sciences USA, 1988,85: 7089 ) 7093
[13] McKendreeW L Jr, Ferl R J. Functional elements of the ArabidopsisAdh promoter include the G-box [J]. Plant Molecular Biol ogy,1992, 19(5) : 859 ) 862
[14] Loake G J,Faktort O,Lamb C J.Combination of H-box [CCTACC(N) 7CT]and G-box (CACGTG) cis elements is necessary for feed-forwardst imulat ion of a chalcone synthase promoter by the phenylpropanoidpathwayintermediate p-coumaric acid [J]. Proceedings of NationalAcademy of Sciences USA, 1992, 89: 9230 ) 9234
[15] Mason H S, DeWald D B, Mullet J E. Ident if ication of a Methyl Jasmonate-Responsive Domain in the Soybean vspB Promot er [J]. ThePlant Cel l, 1993, 5: 241 ) 251
计量
- 文章访问数: 1015
- HTML全文浏览量: 0
- PDF下载量: 573
下载: