甜菜BvMTP11基因的克隆及序列分析
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摘要
在植物中,金属耐受蛋白(Metal tolerant proteins,MTPs)是阳离子转运家族(Cation efflux transporters)重要的成员之一,其在细胞重金属逆境耐受性及离子平衡代谢中发挥着重要的作用。本研究通过RT-PCR方法,在甜菜体内克隆获得了Beta vulgaris metal tolerantce protein 11(BvMTP11)基因的全长序列。该基因CDS全长为1 227 bp,编码了408个氨基酸,分子量(Mw)和等电点(pI)分别为45.935 kDa和4.89,属于相对不稳定蛋白。该基因编码的蛋白有5个跨膜螺旋区,是一种具有阳离子排出域(Cation efflux domain)的膜蛋白,并且有80%的可能性定位于过氧化物酶体膜上。因此可以推测,BvMTP11可能作为甜菜体内重要的转运蛋白参与到重金属逆境胁迫应答网络中,同时,该基因的克隆也为进一步研究其在重金属逆境下的分子机制奠定基础。
Metal tolerance proteins(MTPs) are one of the most important members of Cation efflux transporters in plants. They play an important role in cell heavy metal tolerance and ion homeostasis mechanisms. In this study, Beta vulgaris metal tolerance protein 11(BvMTP11) gene was cloned by RT-PCR in vivo. Its CDS was 1227 bp, which encoded 408 amino acids with molecular weight 45.935 kDa. The theoretical pI was 4.89, and it belonged to a kind of relatively unstable protein. As a membrane protein, BvMTP11 had five membrane spanning regions, and it located in peroxisome membrane with 80% possibility. Thus, it could be concluded that BvMTP11 might function as an essential transport protein involving in sugar beet heavy metal responsive network;meanwhile, cloning will be helpful for further analysis of BvMTP11's molecular mechanism.
Metal tolerance proteins(MTPs) are one of the most important members of Cation efflux transporters in plants. They play an important role in cell heavy metal tolerance and ion homeostasis mechanisms. In this study, Beta vulgaris metal tolerance protein 11(BvMTP11) gene was cloned by RT-PCR in vivo. Its CDS was 1227 bp, which encoded 408 amino acids with molecular weight 45.935 kDa. The theoretical pI was 4.89, and it belonged to a kind of relatively unstable protein. As a membrane protein, BvMTP11 had five membrane spanning regions, and it located in peroxisome membrane with 80% possibility. Thus, it could be concluded that BvMTP11 might function as an essential transport protein involving in sugar beet heavy metal responsive network;meanwhile, cloning will be helpful for further analysis of BvMTP11's molecular mechanism.
引文
[1]Hasan M K,Cheng Y Kanwar M K,et al.Responses of plant proteins to heavy metal stress-a review[J].Frontiers in Plant Science,2017,8:1492.
[2]Ricachenevsky F K,Menguer P K,Sperotto R A,et al.Roles of plant metal tolerance proteins(MTP)in metal storage and potential use in biofortification strategies[J].Frontiers in Plant Science,2013,4:1-16.
[3]Kobae Y,Uemura T,Sato M H,et al.Zinc transporter of Arabidopsis thaliana AtMTP1 is localized to vacuolar membranes and implicated in zinc homeostasis[J].Plant and Cell Physiology,2004,45(12):1749-1758.
[4]Gustin J L,Loureiro M E,Kim D,et al.MTP1-dependent Zn sequestration into shoot vacuoles suggests dual roles in Zn tolerance and accumulation in Zn-hyperaccumulating plants[J].Plant Journal,2009,57(6):1116-1127.
[5]Gustin J L,Zanis M J,Salt D E.Structure and evolution of the plant cation diffusion facilitator family of ion transporters[J].BMCEvolutionary Biology,2011,11:76.
[6]Montanini B,Blaudez D,Jeandroz S,et al.Phylogenetic and functional analysis of the cation diffusion facilitator(CDF)family:Improved signature and prediction of substrate specificity[J].BMC Genomics,2007,8:107.
[7]Wei Y,Fu D.Binding and transport of metal ions at the dimer interface of the Escherichia coli metal transporter YiiP[J].The Journal of Biological Chemistry,2006,281(33):23492-23502.
[8]Arrivault S,Senger T,Kramer U.The Arabidopsis metal tolerance protein AtMTP3 maintains metal homeostasis by mediating Zn exclusion from the shoot under Fe deficiency and Zn oversupply[J].Plant Journal,2006,46:861-879.
[9]Delhaize E,Gruber B,Pittman J,et al.A role for the AtMTP11 gene of Arabidopsis in manganese transport and tolerance[J].Plant Journal,2007,51(2):198-210.
[10]Peiter E,Montanini B,Gobert A,et al.A secretory pathway-localized cation diffusion facilitator confers plant manganese tolerance[J].Proceedings of the National Academy of Sciences of the United States of America,2007,104(20):8532-8537.
[11]Zhang M,Liu B.Identification of a rice metal tolerance protein OsMTP11 as a manganese transporter[J].Plos one,2017,https://doi.org/10.1371/journal.pone.0174987
[12]Liu D,Mao Z,An Z,et al.Enhanced heavy metal tolerance and accumulation by transgenic sugar beets expressing Streptococcus thermophilus StGCS-GS in the presence of Cd,Zn and Cu alone or in combination[J].PLOS ONE,2015,10(6):1-6.
[13]刘大丽,马龙彪,郭爱英,等.镉逆境胁迫下甜菜谷胱甘肽合成酶(BvGS)基因的克隆[J].中国糖料,2017,39(6):23-25.
[14]刘大丽,马龙彪.谷胱甘肽合成酶StGCS-GS对甜菜遗传转化的研究[J].中国糖料,2018,40(5):18-21.
[15]Zhang B J,Zhang X X,Luo L G.The major gene families related to cadmium absorption and transportation in plants[J].Genomics and Applied Biology,2013,32:127-134.
[16]Paulsen I T,Saier M H.A novel family of ubiquitous heavy metal ion transport proteins[J].The Journal of Membrane Biology,1997,156(2):99-103.
[2]Ricachenevsky F K,Menguer P K,Sperotto R A,et al.Roles of plant metal tolerance proteins(MTP)in metal storage and potential use in biofortification strategies[J].Frontiers in Plant Science,2013,4:1-16.
[3]Kobae Y,Uemura T,Sato M H,et al.Zinc transporter of Arabidopsis thaliana AtMTP1 is localized to vacuolar membranes and implicated in zinc homeostasis[J].Plant and Cell Physiology,2004,45(12):1749-1758.
[4]Gustin J L,Loureiro M E,Kim D,et al.MTP1-dependent Zn sequestration into shoot vacuoles suggests dual roles in Zn tolerance and accumulation in Zn-hyperaccumulating plants[J].Plant Journal,2009,57(6):1116-1127.
[5]Gustin J L,Zanis M J,Salt D E.Structure and evolution of the plant cation diffusion facilitator family of ion transporters[J].BMCEvolutionary Biology,2011,11:76.
[6]Montanini B,Blaudez D,Jeandroz S,et al.Phylogenetic and functional analysis of the cation diffusion facilitator(CDF)family:Improved signature and prediction of substrate specificity[J].BMC Genomics,2007,8:107.
[7]Wei Y,Fu D.Binding and transport of metal ions at the dimer interface of the Escherichia coli metal transporter YiiP[J].The Journal of Biological Chemistry,2006,281(33):23492-23502.
[8]Arrivault S,Senger T,Kramer U.The Arabidopsis metal tolerance protein AtMTP3 maintains metal homeostasis by mediating Zn exclusion from the shoot under Fe deficiency and Zn oversupply[J].Plant Journal,2006,46:861-879.
[9]Delhaize E,Gruber B,Pittman J,et al.A role for the AtMTP11 gene of Arabidopsis in manganese transport and tolerance[J].Plant Journal,2007,51(2):198-210.
[10]Peiter E,Montanini B,Gobert A,et al.A secretory pathway-localized cation diffusion facilitator confers plant manganese tolerance[J].Proceedings of the National Academy of Sciences of the United States of America,2007,104(20):8532-8537.
[11]Zhang M,Liu B.Identification of a rice metal tolerance protein OsMTP11 as a manganese transporter[J].Plos one,2017,https://doi.org/10.1371/journal.pone.0174987
[12]Liu D,Mao Z,An Z,et al.Enhanced heavy metal tolerance and accumulation by transgenic sugar beets expressing Streptococcus thermophilus StGCS-GS in the presence of Cd,Zn and Cu alone or in combination[J].PLOS ONE,2015,10(6):1-6.
[13]刘大丽,马龙彪,郭爱英,等.镉逆境胁迫下甜菜谷胱甘肽合成酶(BvGS)基因的克隆[J].中国糖料,2017,39(6):23-25.
[14]刘大丽,马龙彪.谷胱甘肽合成酶StGCS-GS对甜菜遗传转化的研究[J].中国糖料,2018,40(5):18-21.
[15]Zhang B J,Zhang X X,Luo L G.The major gene families related to cadmium absorption and transportation in plants[J].Genomics and Applied Biology,2013,32:127-134.
[16]Paulsen I T,Saier M H.A novel family of ubiquitous heavy metal ion transport proteins[J].The Journal of Membrane Biology,1997,156(2):99-103.