铁观音茶树转录因子TCP4基因的克隆与序列分析
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摘要
以铁观音茶树鲜叶为研究材料,采用RACE-PCR的方法,根据已克隆出的植物TCP转录因子序列,设计特异引物,克隆获得TCP4基因的cDNA全长序列,命名为CsTCP4,其基因全长c DNA为1264 bp,包含一个完整的795 bp开放阅读框,编码264个氨基酸。生物信息学分析表明,该基因编码的蛋白质属于不稳定亲水性蛋白;通过同源进化树分析得出该铁观音茶树基因CsTCP4与草莓、桃树等的TCP基因同源性较高,属于TCP家族的CIN分支,且其编码的蛋白质具有TCP基因家族共有的结构特征,即bHLH结构域,说明CsTCP4在叶片生长与发育方面起到重要的调控作用,对调控铁观音茶树叶片性状分化尤为关键。
The full-length c DNA of TCP4 gene was cloned from Tieguanyin tea( Camellia sinensis) by using RACE-PCR according to the cloned plant TCP transcription factors sequences. Named as CsTCP4,it had a full length of 1,264 bp with intact ORFs of 795 bp and encoding 264 amino acids. Various bioinformatic analyses showed it to be homologous to the TCP genes of Fragaria vesca subsp. vesca and Prunus persica,and its encoded protein with the structural characteristics of TCP family. The protein structure was basic-Helix-Loop-Helix( b HLH) and belonged to the CIN branch of TCP family. The TCP4 gene could be used as the target gene of miR329,which involves mainly in the cell differentiation and development of a plant,especially in the regulation of leaf growth and development. Consequently,CsTCP4 could be a crucial factor associated with the formation and characteristics of Tieguanyin tea leaves.
The full-length c DNA of TCP4 gene was cloned from Tieguanyin tea( Camellia sinensis) by using RACE-PCR according to the cloned plant TCP transcription factors sequences. Named as CsTCP4,it had a full length of 1,264 bp with intact ORFs of 795 bp and encoding 264 amino acids. Various bioinformatic analyses showed it to be homologous to the TCP genes of Fragaria vesca subsp. vesca and Prunus persica,and its encoded protein with the structural characteristics of TCP family. The protein structure was basic-Helix-Loop-Helix( b HLH) and belonged to the CIN branch of TCP family. The TCP4 gene could be used as the target gene of miR329,which involves mainly in the cell differentiation and development of a plant,especially in the regulation of leaf growth and development. Consequently,CsTCP4 could be a crucial factor associated with the formation and characteristics of Tieguanyin tea leaves.
引文
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[2]安家兴.拟南芥TCP11调控维管束的发育[D].兰州:兰州大学,2012.
[3]张春雷.拟南芥TCP15、TCP22基因功能的研究[D].兰州:兰州大学,2012.
[4]王玉红,徐立华,李军.玉米TCP家族基因生物信息学鉴定与分析[J].山东农业科学,2014(4):1-6.
[5]朱姗姗.水稻转录因子PCF7的功能研究[D].重庆:重庆大学,2015.
[6]刘伟娜,韩建民,董金皋,等.棉花TCP家族转录因子基因GhTCP1的克隆与表达分析[J].棉花学报,2010,22(5):409-414.
[7]李燕,姚金波,陈伟,等.雷蒙德氏棉和亚洲棉TCP基因家族的生物信息学分析[J].棉花学报,2016,28(5):434-442.
[8]张士刚.苹果TCP转录因子家族生物信息学分析[J].山东农业科学,2014,46(5):12-17.
[9]袁伟,顾掌根,褚伟雄,等.黄瓜转录因子TCP基因家族的鉴定及结构分析[J].分子植物育种,2014,12(2):287-295.
[10]冀志蕊,谷彦冰,董庆龙,等.葡萄TCP基因家族全基因组鉴定和分析[J].基因组学与应用生物学,2015,34(10):2194-2199.
[11]时丕彪.西瓜TCP转录因子的鉴定及功能分析[D].杭州:浙江大学,2016.
[12]邹世慧,王会平,余勇,等.矮牵牛ECE支TCP基因的克隆及表达分析[J].园艺学报,2013,40(2):307-316.
[13]邹世慧.矮牵牛ECE支TCP基因的克隆、表达模式、序列变异及PhTCP1的超量表达研究[D].重庆:西南大学,2012.
[14]王会平.矮牵牛PhTCP2基因的克隆、表达分析及超量表达研究[D].重庆:西南大学,2013.
[15]余勇.矮牵牛PhTCP3、PhTCP4基因功能的初步研究[D].重庆:西南大学,2014.
[16]周瑜.拟南芥转录因子TCP17参与光信号转导和生长素合成[D].兰州:兰州大学,2013.
[17]Takeda T,Amano K,Ohto M A,et al.RNA interference of the Arabidopsis putative transcription factor TCP16 gene results in abortion of early pollen development[J].Plant Molecular Biology,2006,61(1/2):165-177.
[18]Li C,Potuschak T,Colon-Carmona A,et al.Arabidopsis TCP20links regulation of growth and cell division control pathways[J].Proceedings of the National Academy of Sciences of the United States of America,2005,102(36):12978-12983.
[19]Aguilar-Martinez J A,Sinha N.Analysis of the role of Arabidopsis class I TCP genes At TCP7 At TCP8 At TCP22 and At TCP23 in leaf development.Front Plant Sci,,2013,4(1):1-13.
[20]陈志丹.铁观音茶树种性分化的分子鉴定及差异表达基因分离研究[D].福州:福建农林大学,2013.
[21]程国山.茶树CSAG基因克隆及AG基因系统进化分析[D].西安:陕西师范大学,2014.
[22]李娟.安吉白茶高氨基酸性状相关基因的全长c DNA克隆及功能的初步研究[D].长沙:湖南农业大学,2011.
[23]胡波.茶树成花相关基因的c DNA克隆及生物信息学分析[D].福州:福建农林大学,2010.
[24]何志敏.拟南芥转录因子TCP2与CRY1相互作用的遗传学与生化分析[D].长沙:湖南大学,2016.
[25]刘丽娟,高辉.TCP家族基因研究进展[J].生物技术通报,2016,32(9):14-22.
[26]许梦楠.TCP基因调控黄瓜卷须发育机理的初探[D].北京:中国农业科学院,2015.
[27]Nath U,Crawford B C,Carpenter R,et al.Genetic control of surface curvature[J].Science,2003,299(5611):1404-1407.