Cloning and Function Analysis of a Flower-specific Expression Promoter PCHS

Abstract

Abstract: A polymerase chain reaction（PCR）amplification was used to isolate a flower-specific expression promoter PCHS from Arabidopsis genomic DNA. Sequence analysis revealed that PCHS was 531 bp in total length and shared significant nucleotides identity 99% with GenBank databases, which contained four flower-specific expression regulatory elements. Intermediate vector pPCHS was constructed by inserting the promoter to the upstream of GUS in pB1121, where the original CaMV35S promoter was replaced by the PCHS promoter. The flower, stem, leaf ,and root of petunia hybrida were transformed via Agrobacterium tumefaciens. Histochemical localization of GUS activity indicated that the PCHS promoter could confer a flower-specific expression pattern to GUS.

Li Gang, Tao Meiying, Jia Caihong,Zhang Jianbin, Xu Biyu Jin Zhiqiang. Cloning and Function Analysis of a Flower-specific Expression Promoter PCHS[J]. Chinese Agricultural Science Bulletin, 2008, 24(4): 89-93.

References

[1] 何生根. 切花品质的生理生化基础. 植物生理学通讯, 1997, 33(1): 66- 70.
[2] 曾海涛, 王义琴, 陈英, 等. 花卉花色基因工程的研究现状及存在问题. 中国生物工程杂志, 2004, 24(6): 54- 57.
[3] Gawel N J, Jarret R L. A modified CTAB DNA extraction procedure for Mesa and Ipomoea[J]. Plant Molecular Biology Reporter, 1991, 9: 262- 266.
[4] Koch,M.A., Weisshaar,B., Kroymann,J., et a1. Comparative genomics and regulatory evolution: conservation and
function of the Chs and Apetala3 promoters. Molecular Biology and Evolution, 2001, 18 (10): 1882- 1891.
[5] 黄培堂译. 分子克隆实验指南. 北京: 科学出版社, 2002: 1116- 1125.
[6] 王关林, 方宏箔主编. 植物基因工程原理与技术. 北京: 科学出版社, 1998: 428- 430.
[7] Schulze-Lefert P, Dang1 J L, Becker-Andre M, et a1. Inducible in vivo DNA footprints define sequences necessary for UV light activation of the parsley chalcone synthase gene. EMBO, 1989, 8: 651- 656.
[8] Staiger D. Kaulen H, Sche11 J. A CACGTG motif of the Antirrhinum majus chalcone recoginized by an evolutionary synthase conserved promoter is nuclear protein[J]. Proc Nat1 Acad Sci USA, 1989, 86: 6930- 6934.
[9] Guiliano G, Pichersky E, Ma1ik V S. An evolutionary conserved protein binding sequence upstream of a plant light-regulated gene[J]. Proc Nat1 Acad Sci USA, 1988, 85: 7089- 7093
[10] Hamilton D A., Schwarz Y H., Mascarenhas J P. A monocot pollen-specific promoter contains separable pollen-specific and quantitative elements[J]. Plant Molecular Biology, 1998, 38: 663- 669.
[11] Mitsuda N, Takeyasu K, Sato M H. Pollen-specific regulation of vacuolar H＋-PPase expression by multiple cis -acting elements[J]. Plant Molecular Biology, 2001, 46: 185- 192.
[12] Morris S H., Adley C C. Irish public perceptions and attitudes to modern biotechnology: an overview with a focus on GM foods[J]. Trends in Biotechnology, 2001, 19(2): 43- 48.
[13] Simth N. Seeds of opportunity an assessment of the benefits. safety and oversight of plant enomics and agricultural biotechnology[J]. Committee Print. 2000: 106- 111.
[14] Kagaya Y., Nakamura H., Hidaka S., Ejiri Sl. and Tsutsumi KI. The promoter from the rice nuclear gene encoding chloroplast aldolase confers mesophyll-specific and light regulated expression in transgenic tobacco[J]. Molecular and General Genetics, 1995, 248 (6): 668- 674.
[15] Nomura M., Katayama K., Nishimura A. The promoter of rbcS in a C3 plant (rice) directs organ specific, light dependent expression in a C4 plant (maize), but does not confer bundle sheath cell specific expression[J]. Plant Molecular Biology, 2000, 44 (1): 99- 106.
[16] 赵恢武, 陈杨坚, 胡莺雷, 高音和, 林忠平. 干旱诱导性启动子驱动的海藻糖-6-磷酸合酶基因载体的构建及转基因烟草的耐旱性[J]. 植物学报, 2000, 42(6): 616- 619.
[17] Faktor O,kooter J M, Dixon R A, et al. Functional dissection of a bean chalcone svnthase gene promoter in transgenic: tobacco plants reveals sequence motifs essential for floral caprassion. PMB, 1996,32: 849- 859.
[18] Seung H Y, Yube Y, Nozomu K, Tomonobu K,Hiroshi S. Promoter analysis of tbzF, a gene encoding a bZIP-type transcription factor, reveals distinct variation in cis-regions responsible for transcriptional activation between senescing leaves and flower buds in tobacco plants[J]. Plant Science, 2002, 162: 973- 980.
[19] Weterings, J. Schrauwen, G. Wullems, D. Twell. Functional dissection of the promoter of the pollen-specific gene NTP303 reveals a novel pollen-specific, and conserved cis –regulatory element[J]. Plant Journal, 1995, 8: 55- 63.
[20] Qian Y Q, Ma K P. Progress in the study on genetically modified organisms and the impact of its release on the environment. Acta Ecol Sin, 1998, 18(1): 1- 9.
[21] Hancock J F, Grumet R, Hokanson S C. The opportunity for escape of engineered gene from transgenic crops. Hortscience, 1996, 31(7): 1080- 1085.
[22] Chevre A M, Eber E, Renard M. Transgenic rape and environmental hazards. Biofutur, 1998, 172: 44- 48.