土人参Actin基因片段的克隆及序列分析
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摘要
为土人参功能基因的表达与调控提供内参基因,根据已知植物Actin基因的保守区设计简并性引物,采用RT-PCR方法扩增得到土人参的Actin基因片段,将该片段连接于p GEM-T载体,测序获得一段大小为598 bp的基因片段,该片段编码198个氨基酸。通过生物信息学软件分析,该序列与其他植物Actin基因的cDNA序列同源性均在85%以上,氨基酸序列的同源性达到86%以上,表明试验克隆所得基因片段为土人参Actin基因片段。将该Actin基因片段命名为TpActin1,并登陆在Gena Bank,登录号为MH333039。
In order to provide reference gene for studying the expression pattern of functional genes in Talinum paniculatum,the cDNA fragment of Actin from Talinum paniculatum;was amplified by RT-PCR,by adopting degenerated primers designed according to the conserved region of the Actin genes which were submitted to NCBI database. The cDNA fragment was inserted into the p GEM-T vector and sequenced. The cDNA sequence was analyzed by online bioinformatics software. Our result showed that this actin cDNA includes 598 base pairs,encoding a partial actin with 198 amino acids. Sequence alignments showed that it shares over 85% nucleotide sequence identity and 86% amino acid sequence identity with other plant actins. This study indicated that a new actin cDNA fragment was isolated from Talinum paniculatum. This cDNA was named TpActin1 and submitted to GenBank (accession number: MH33309).
In order to provide reference gene for studying the expression pattern of functional genes in Talinum paniculatum,the cDNA fragment of Actin from Talinum paniculatum;was amplified by RT-PCR,by adopting degenerated primers designed according to the conserved region of the Actin genes which were submitted to NCBI database. The cDNA fragment was inserted into the p GEM-T vector and sequenced. The cDNA sequence was analyzed by online bioinformatics software. Our result showed that this actin cDNA includes 598 base pairs,encoding a partial actin with 198 amino acids. Sequence alignments showed that it shares over 85% nucleotide sequence identity and 86% amino acid sequence identity with other plant actins. This study indicated that a new actin cDNA fragment was isolated from Talinum paniculatum. This cDNA was named TpActin1 and submitted to GenBank (accession number: MH33309).
引文
[1]郭艳,张美,夏快飞,等.厚藤Actin基因片段的克隆及序列分析[J].氨基酸和生物资源,2016,38(3):59-62.
[2]Mccurdy D W,Kovar D R,Staiger C J.Actin and actin-binding proteins in higher plants[J].Protoplasma,2001,215(1-4):89-104.
[3]郭艳,夏快飞,张美,等.滨海适生植物草海桐Actin基因片段的克隆及序列分析[J].生物技术世界,2016(5):39-40.
[4]崔力文,郑婷,张克坤,等.葡萄Actin基因家族的鉴定及进化和表达分析[J].植物资源与环境学报,2017,26(3):1-10.
[5]孔卫青,杨金宏.桑树肌动蛋白Actin基因全长序列的克隆与分析[J].广西植物,2012,32 (3):362-366.
[6]郭欢,崔彦农,吴凡,等.盐生植物四翅滨藜Actin基因片段的克隆及表达分析[J].草业科学,2017,34(3):515-522.
[7]马清,周向睿,伍国强,等.盐生植物碱蓬Actin基因片段的克隆及序列分析[J].生物技术,2009,19(1):1-3.
[8]Muthugapatti K K,Elizabeth C M,Richard B M.Functional nonequivalency of Actin isovariants in Arabidopsis[J].Molecular Biology of the Cell,2002,13(1):251.
[9]中国科学院中国植物志委员会.中国植物志.第二十六卷,被子植物门,双子叶植物纲,紫茉莉科 马齿苋科 商陆科 落葵科 番杏科 石竹科[M].北京:科学出版社,1996.
[10]王翔,林梦瑶,黄锁义,等.土人参研究的新进展[J].微量元素与健康研究,2017,34(1):75-77.
[11]刘晓珍,汤艳姬,陈斯钊,等.土人参黄酮类化合物的提取及其抗氧化性[J].贵州农业科学,2012,40(11):192-195.
[12]黄礼德,郭立强,潘廷啟,等.正交设计优化土人参多糖的超声提取工艺[J].中国酿造,2012,31(3):105-107.
[13]Dos R L F C,Cerdeira C D,De P B F,et al.Chemical characterization and evaluation of antibacterial,antifungal,antimycobacterial,and cytotoxic activities of Talinum paniculatum[J].Revista Do Instituto De Medicina Tropical De S?o Paulo,2015,57(5):397-405.
[14]文全泰,潘廷啟,黄礼德,等.土人参提取液对D-半乳糖所致衰老小鼠的抗氧化作用研究[J].天然产物研究与开发,2014,26(5):662-665.
[15]Zhu X,Zeng X,Huang B,et al.Actin is closely associated with RNA polymerase II and involved in activation of gene transcription[J].Biochem Biophys Res Commun,2004,321(3):623-630.
[16]Kozuka J,Yokota H,Arai Y,et al.Dynamic polymorphism of actin as activation mechanism for cell motility[J].Biosystems,2007,88(3):273-282.
[17]Papakonstanti E A,Stournaras C.Cell responses regulated by early reorganization of actin cytoskeleton.[J].Febs Letters,2008,582 (14):2120-2127.
[18]Chang T J,Juan C C,Yin P H,et al.Upregulation of beta-actin,cyclophilin and GAPDH in N1S1 rat hepatoma.[J].Oncology Reports,1998,5(2):469.
[19]Calvo E,Rubiano C,Vargas A,et al.Expression of housekeeping genes during the asexual cell cycle of Plasmodium falciparum[J].Parasitology Research,2002,88(3):267-271.
[20]Mcdowell J M,Huang S,Mckinney E C,et al.Structure and evolution of the actin gene family in Arabidopsis thaliana.[J].Genetics,1996,142(2):587.
[21]许娟,罗兴录.木薯Actin基因片段的克隆及序列分析[J].生物技术通报,2011(6):65-70.
[22]伍国强,席杰军,包爱科,等.多浆旱生植物霸王Actin基因片段的克隆及序列分析[J].生物技术通报,2008(2):101-104.
[23]张灵,段慧荣,李静,等.旱生植物梭梭Actin基因片段的克隆及表达模式分析[J].基因组学与应用生物学,2014,33(3):598-603.
[2]Mccurdy D W,Kovar D R,Staiger C J.Actin and actin-binding proteins in higher plants[J].Protoplasma,2001,215(1-4):89-104.
[3]郭艳,夏快飞,张美,等.滨海适生植物草海桐Actin基因片段的克隆及序列分析[J].生物技术世界,2016(5):39-40.
[4]崔力文,郑婷,张克坤,等.葡萄Actin基因家族的鉴定及进化和表达分析[J].植物资源与环境学报,2017,26(3):1-10.
[5]孔卫青,杨金宏.桑树肌动蛋白Actin基因全长序列的克隆与分析[J].广西植物,2012,32 (3):362-366.
[6]郭欢,崔彦农,吴凡,等.盐生植物四翅滨藜Actin基因片段的克隆及表达分析[J].草业科学,2017,34(3):515-522.
[7]马清,周向睿,伍国强,等.盐生植物碱蓬Actin基因片段的克隆及序列分析[J].生物技术,2009,19(1):1-3.
[8]Muthugapatti K K,Elizabeth C M,Richard B M.Functional nonequivalency of Actin isovariants in Arabidopsis[J].Molecular Biology of the Cell,2002,13(1):251.
[9]中国科学院中国植物志委员会.中国植物志.第二十六卷,被子植物门,双子叶植物纲,紫茉莉科 马齿苋科 商陆科 落葵科 番杏科 石竹科[M].北京:科学出版社,1996.
[10]王翔,林梦瑶,黄锁义,等.土人参研究的新进展[J].微量元素与健康研究,2017,34(1):75-77.
[11]刘晓珍,汤艳姬,陈斯钊,等.土人参黄酮类化合物的提取及其抗氧化性[J].贵州农业科学,2012,40(11):192-195.
[12]黄礼德,郭立强,潘廷啟,等.正交设计优化土人参多糖的超声提取工艺[J].中国酿造,2012,31(3):105-107.
[13]Dos R L F C,Cerdeira C D,De P B F,et al.Chemical characterization and evaluation of antibacterial,antifungal,antimycobacterial,and cytotoxic activities of Talinum paniculatum[J].Revista Do Instituto De Medicina Tropical De S?o Paulo,2015,57(5):397-405.
[14]文全泰,潘廷啟,黄礼德,等.土人参提取液对D-半乳糖所致衰老小鼠的抗氧化作用研究[J].天然产物研究与开发,2014,26(5):662-665.
[15]Zhu X,Zeng X,Huang B,et al.Actin is closely associated with RNA polymerase II and involved in activation of gene transcription[J].Biochem Biophys Res Commun,2004,321(3):623-630.
[16]Kozuka J,Yokota H,Arai Y,et al.Dynamic polymorphism of actin as activation mechanism for cell motility[J].Biosystems,2007,88(3):273-282.
[17]Papakonstanti E A,Stournaras C.Cell responses regulated by early reorganization of actin cytoskeleton.[J].Febs Letters,2008,582 (14):2120-2127.
[18]Chang T J,Juan C C,Yin P H,et al.Upregulation of beta-actin,cyclophilin and GAPDH in N1S1 rat hepatoma.[J].Oncology Reports,1998,5(2):469.
[19]Calvo E,Rubiano C,Vargas A,et al.Expression of housekeeping genes during the asexual cell cycle of Plasmodium falciparum[J].Parasitology Research,2002,88(3):267-271.
[20]Mcdowell J M,Huang S,Mckinney E C,et al.Structure and evolution of the actin gene family in Arabidopsis thaliana.[J].Genetics,1996,142(2):587.
[21]许娟,罗兴录.木薯Actin基因片段的克隆及序列分析[J].生物技术通报,2011(6):65-70.
[22]伍国强,席杰军,包爱科,等.多浆旱生植物霸王Actin基因片段的克隆及序列分析[J].生物技术通报,2008(2):101-104.
[23]张灵,段慧荣,李静,等.旱生植物梭梭Actin基因片段的克隆及表达模式分析[J].基因组学与应用生物学,2014,33(3):598-603.