油菜转录因子BnZFP基因的分离及其功能初步分析

摘要/Abstract

摘要： 为克隆油菜ZFP类转录因子基因，初步阐明其功能。采用RT-PCR的方法克隆基因和体内检测；花沾法转化拟南芥，Northern blotting检测转基因后的表达。从甘蓝型油菜中分离了一个新的ZFP类转录因子，它编码377个氨基酸，与拟南芥AtZFP基因(At2g29660)的碱基序列相似性最高，为76.2%，命名为BnZFP。该基因在父本、母本和F1代不同发育时期中均能较稳定的表达；该基因转化拟南芥，对获得的转BnZFP基因拟南芥进行了Northern Blotting分析，表明该基因在拟南芥中得到过表达；与野生型拟南芥相比较，过表达BnZFP的转基因拟南芥在萌发、营养生长及生殖生长过程中没有表现出明显的表型变化，该基因的功能有待进一步研究。

关键词: 山羊, 山羊, 表皮干细胞, 分离纯化

Abstract: To clone a new transcription factor from Brassica napus, and study its function. Semi-quantitative RT-PCR, Transformation, Northern-blotting. BnZFP was isolated from Brassica.napus, which constituted 377 amino acids. BnZFP had the highest similarity with AtZFP at nucleotide and amino acids level. Semi-quantitative RT-PCR showed BnZFP was steadily expressed throughout the different developmental stages between F1 and parental lines. BnZFP was constructed to pBI121 vector and this expression cassette was introduced into wild Arabidopsis thaliana. The transgenic plants were analysed by Northern blotting. Compared with wild Arabidopsis thaliana, over-expression of BnZFP in plants didn’t show any visible changes in seed germination and post-germination growth, further study should be carried out to study the gene’s function.

中图分类号:

Q786

张兰,,王磊,,涂金星,范云六,. 油菜转录因子BnZFP基因的分离及其功能初步分析[J]. 中国农学通报, 2007, 23(2): 45-045.

Zhang Lan,, Wang Lei,, Tu Jinxing, Fan Yunliu,. Cloning and Preliminary Characterization of the BnZFP Gene from Brassica napus[J]. Chinese Agricultural Science Bulletin, 2007, 23(2): 45-045.

参考文献

[1] Takatsuji H. Zinc-finger transcription factors in plants[J]. Cell Mol Life Sci, 1998, 54: 582-596.
[2] Rowman J L, Sakai H, Jack T, et al. SUPERMAN, a regulator of floral homeotic genes in Arabidopsis[J]. Development, 1992,114: 599-615.
[3] Sakai H, Medrano L J, Meyerowitz E M. Role of SUPERMAN in maintaining Arabidopsis floral whorl boundaries[J]. Nature, 1995, 378: 199-203.
[4] Yun J Y, Weigel D, Lee I. Ecotopic expression of SUPERMAN suppresses development of petals and stamens[J]. Plant Cell Physiol, 2002, 43: 52-57.
[5] Tague B W, Gallant P, Goodman H M. Expression analysis of an Arabidopsis C2H2 zinc finger protein[J]. Plant Mol Biol, 1997, 32: 785-796.
[6] Chrispeels H E, Oettinger H, Janvier N, et al. At-ZFP1, encoding Arabidopsis thaliana C2H2 zinc-finger 1, is expressed downstream of photomorphogenic activation[J]. Plant Mol Biol, 2000, 42: 279-290.
[7] Luo M, Bilodeau P, Koltunow A, et al. Genes controlling fertilization-independent seed development in Arabidopsis thaliana[J]. Pro Natl Acad Sci USA, 1999, 96: 296-301.
[8] Prigge M J, Wagner D R. The Arabidopsis SERRATE gene encodes a zinc-finger protein required for normal shoot development[J]. Plant Cell, 2001,13:1263-1279.
[9] Kubo K, Sakamoto A, Kobayashi A, et al. Cys2/His2 zinc-finger protein family of petunia: evolution and general mechanism of target-sequence tecognition[J]. Nucleic Acids Res, 1998, 26:608-615.
[10] Takatsuji H, Mori M, Benfey P N, et al. Characterization of a zinc finger DNA-binding protein expressed specifically in petunia petals and seedlings[J]. EMBO J, 1992,11:241-249.
[11] Van Der Krol A R, Van Poecke R M, Vorst O F, et al. Development and wound-, cold-,desiccation-, ultraviolet-B-stress-induced modulations in the expression of the petunia zinc finger transcription factor gene ZPT2-2[J]. Plant Physiol, 1999, 121: 1153-1162.
[12] Kubo K, Kanno Y, Nishino T, et al. Zinc-finger genes that specifically express in pistil secretory tissues of petunia[J]. Plant Cell Physiol, 2000, 41: 377-382.
[13] Sugano S, Kaminaks H, Rybks Z, et al. Stress-responsive Zinc finger gene ZPT2-3 plays a role in drought tolerance in petunia[J]. Plant J, 2003, 36: 830-841.
[14] Sakamoto A, Minami M, Huh G H, et al. The putative zinc-finger protein WZF1 interacts with a cis-acting element of wheat histone genes[J]. Eur J Biochem, 1993, 217:1049-1056.
[15] 黄骥,张红生,曹雅君,等.一个新的水稻C2H2型锌指蛋白cDNA的克隆与序列分析[J].南京农业大学学报, 2002, 25(2): 110-112.
[16] Benjamin L. Gene Ⅷ. Oxford University Press[D]: 2000,654-656.
[17] Michael J P, Wagner D R. The Arabidopsis SERRATE gene encodes a zinc-finger protein required for normal shoot development[J]. Plant Cell, 2001, 13: 1263-1279.
[18] Kobayashi A, Sakmoto A, Kubo K, et al. Seven zinc-finger transcription factors are expressed sequentially during the development of anthers in petunia[J]. Plant J, 1998, 13(4): 571-576.
[19] 王东,杨金水.棉花类耐盐锌指蛋白基因的克隆与结构分析[J].复旦学报(自然科学版), 2002, 41(4): 42-46.