WEI Wen-wen,WANG Pi-wu,FU Yong-ping,et al.Cloning and Identification of the Seedcoat Specific Promoter from Soybean[J].Soybean Science,2010,29(03):390-393.[doi:10.11861/j.issn.1000-9841.2010.03.0390]
大豆种皮特异性启动子的克隆及功能分析
- Title:
- Cloning and Identification of the Seedcoat Specific Promoter from Soybean
- 文章编号:
- 1000-9841（2010）03-0390-04
- 分类号:
- S565.1
- 文献标志码:
- A
- 摘要:
- 采用PCR技术从大豆品种吉农17基因组DNA中分离得到大豆种皮特异性启动子片段SCSP,长度约为1268 bp。序列分析表明SCSP与报道序列同源性达97%以上。将其与GUS基因融合,构建植物表达载体pSCSP-GUS后,转化到EHA105根癌农杆菌中,通过根癌农杆菌(Agrobacterium tumefaciens)介导法转化烟草(Nicotiana tabacum)NC89。转基因植株的GUS活性检测表明,仅能在种皮检测到GUS活性,而在茎、叶和花等其它组织中都未检测到GUS活性,证实SCSP片段具有种皮特异性表达功能。
- Abstract:
- The promoter region of Seedcoat-specific gene was isolated from the genomic DNA of soybean Jilin 17 by PCR method, and obtained the promoter fragment SCSP, the length of the clone sequence is 1268bp. Sequence analysis indicated that this fragment had 97% homology compared with the reported promoters. The cloned promoter was fused to the GUS reporter gene to construct plant expression vector pSCSP-GUS,which was transferred into tobacco(Nicotiana tabacum)NC89 by Agrobacterium tumefaciens-mediated method and severa1transformed plants were obtained.GUS activity assays of different soybean organs indicated that expression of GUS was active only in seedcoat,which suggests the SCSP gene is seedcoat-specific promoter.
参考文献/References:
[1]李桂兰,王文颇,臧少先,等. 拟南芥黑芥子酶基因根特异性表达启动子的克隆[J].河北农业大学学报, 2005, 28 (1) : 52-55.(Li G L,Wang W P, Zang S X, et al. Cloning of root specif ic expression promoter of myrosinase ( pyk10 ) gene from Arabidopsis thaliana[J]. Journal of Agricultural University of Hebei, 2005, 28(1): 52-55.) [2]Romero C ,Belles J M ,Vaya J L ,et al. Expression of the yeast trehalose-6-phosphate synthase gene in transgenic tobacco plants: pleiot ropic phenotypes include drought tolerance[J]. Planta,1997,201:293-297. [3]Eaglesham A , Pueppke S G, Hardy R W F，et al. Gentically modifiedfood and the consumer. NABC Report 13. New York: National Agricultural Biotechnology Council,2001. [4]张春晓,王文棋,蒋湘宁,等. 植物基因启动子研究进展[J].遗传学报, 2004,31(12):130-139. ( Zhang C X, Wang W Q, Jiang X N, et al. Review on plant gene promoters[J]. Acta Genetica Sinica, 2004,31(12):130-139.) [5]Shirsat A,Wilfword N, Croy R, et al. Sequences responsible for the tissue specific promoter activity of a pea legumin gene in tobacco[J].Molecular and General Genetics, 1989, 215: 326-331. [6]肖娜,武玉花.油菜种子特异表达启动子pNapB和pFAE1的结构与功能比较研究[J].中国油料作物学报,2008,30 (1):1-9. ( Xiao N, WU Y H. Structural and function analysis of two seed-specific promoters: pNapB and pFAE1from B rassica napus[J]. Chinese Journal of Oil Crop Sciences 2008,30 (1):1-9.) [7]王关林,方宏筠. 植物基因工程[M]. 北京:科学出版社, 2002:744. (Wang G L, Fang H J. Plant genetic engineering[M]. Beijing: Science Press, 2002: 744.) [8]Hofen R,Willmitzer L. Storage of competent cells for Agrobacterium transformation[J]. Nucleic Acids Research,1988,16:9877. [9]Horsch R B, Fry J E, Hoffman N L.A simple and general method for transferring gene into plant[J]. Science, 1985,227:1229-1231. [10]Jefferson R A. Assaying chimeric genes in plants: The GUS genefusion system[J]. Plant Molecular Biology Reporter,1987,5(4):387-405. [11]付永平，周海涛，王丕武.大豆种子特异性启动子的克隆及功能分析[J].西北农林科技大学学报，2009，37（12）：105-118.（Fu Y P, Zhou H T, Wang P W. Cloning and identification of the seed specific promoter from soybean[J]. Journal of Northwest A & F University, 2009, 37（12）: 105-118.) [12]Nishiuchi T ,Shinshi H ,Suzuki K. Rapid and transient activation of transcription of the ERF3 gene by woun- ding in tobacco leaves :Possible involvement of NtWRKYs and autorepression [J]. Journal of Biological Chemistry,2004,279: 55355-55361.
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备注/Memo
基金项目：国家自然科学基金资助项目(30971805)； 教育部博士点基金资助项目（20070193005）。