AtMGT9基因T-DNA插入突变体表型分析

利用功能互补, 对拟南芥AtMGT9基因的Mg2+转运功能进行了研究。同时对AtMGT9的两个T-DNA插入突变体株系(mgt9-1,mgt9-2)进行表型分析, mgt9-1只能得到杂合体植株, 有花粉败育现象, mgt9-2有纯合体, 但没有可见表型。基因序列分析表明, mgt9-1中T-DNA插入位点在起始密码子后, mgt9-2中T-DNA插入位点在靠近C端第2个跨膜区中间位置。功能互补和63Ni2+示踪实验证明截短的mgt9-2蛋白能够互补细菌镁离子转运突变体MM281, 具有全序列基因一样的转运特性, 而mgt9-1不具有转运功能, 是一个功能缺失突变体。这为进一步研究AtMGT9蛋白的结构与功能的关系奠定了基础。

Abstract

Functional complementation showed that AtMGT9 functions as a Mg2+ transporter. Compared with the two T-DNA insertional mutant mgt9-1 and mgt9-2 in AtMGT9, there are only heterozygote plants of mgt9-1 and that have pollen abortion phenomenon. The mgt9-2 are homozygous, but no visible phenotype. The T-DNA insertion point of mgt9-1 is in the initiation codon position,but the T-DNA insertion point of mgt9-2 is in the stop codon before the second transmembrane. Functional complementation and the 63Ni2+ tracer analysis shows that mgt9-2 functions as a Mg2+ transporter. This laid the foundation for the further study of the relationship of AtMGT9 protein structure and function.

参考文献

[1] SMITH R L, MAGUIRE M E. Microbial Magnesium Transport: Nnusual Transports Searching for Identity［J］. Mol Microbio, 1998, 28(2): 217-226.

[2] HORLITZ M, KLAFF P. Gene-specific Trans-regulatory Functions of Magnesium for Chloroplast mRNA Stability in Higher Plants［J］. J Biol Chem, 2000, 275(45): 35638-35645.

[3] COLIN W M, RICHARD C G. Overexpression of the Saccharnoy-ces Cerevisiae Magnesium Transport System Confers Resistance to Aluminumion［J］. J Biol Chem, 1998, 273(3): 1727-1732.

[4] LI L,TUTONE A F, DRUMMOND R S M,et al.A Novel Family of Magnesium Transport Genes in Arabidopsis［J］. The Plant Cell, 2001, 13(12): 2761-2775.

[5] SILVER S, KRALOVIC M L. Manganese Accumulation by Escherichia coli: Evidence for a Specific Transport System［J］. Biochem Biophys Res Commun, 1969, 34: 640-645.

[6] BUI D M, GREGAN J, JAROSCH E,et al.The Bacterial Magnesium Transporter CorA can Functionally Substitute for Its Putative Homologue Mrs2p in the Yeast Inner Mitochondrial Membrane［J］. J Biol Chem, 1999, 274(29): 20438-20443.

[7] MAO DAN-DAN, TIAN LIAN-FU, LI LE-GONG, et al. AtMGT7: An Arabidopsis Gene Encoding a Low-Affinity Magnesium Transporter ［J］. JIPB, 2008, 50(12): 1530-1538.

[8] SCHOCK I, GREGAN J, STEINHAUSER S, et al. A Member of a Novel Arabidopsis Thaliana Gene Family of Candidate Mg2+ ion Transporters Complements a Yeast Mitochondrial Group Intron-splicing Mutant ［J］.The Plant Journal, 2000, 24(4): 489-501.

[9] CLOUGH S J, BENT A F. Floral Dip: a Simplified Method for Agrobacterium-mediated Transformation of Arabidopsis thaliana［J］. Plant J, 1998, 16: 735-743.

[10] CHEN JIAN, LI LE-GONG, LIU ZHEN-HUA, et al. Magnesium Transporter AtMGT9 is Essential for Pollen Development in Arabidopsis［J］. Cell Res, 2009, 19: 887-898.

[11] SZEGEDY M A,MAGUIRE M E. The CorA Mg2+ Transport Protein of Salmonella typhimurium Mutagenesis of Conserved Residues in the Second Membrane Domain［J］. J Biol Chem, 1999, 274: 36973-36979.

[12] SMITH R L, SZEGEDY M A, KUCHARSKI L M,et al. The CorA Mg2+ Transport Protein of Salmonella typhimurium: Mutagenesis of Conserved Residues in the Third Membrane Domain Identifies a Mg2+ Pore［J］. J Biol Chem, 1998, 273: 28663-28669.

史佳, 李健, 程伟, 陈键, 田连福, 朱俊林, 李东屏. AtMGT9基因T-DNA插入突变体表型分析[J]. 激光生物学报, 2010, 19(6): 778. SHI Jia, LI-jian, CHENG Wei, CHEN Jian, TIAN Lian-fu, ZHU Jun-lin, LI Dong-ping. Phenotypic Analysis of the AtMGT9 T-DNA Insertional Mutants[J]. Acta Laser Biology Sinica, 2010, 19(6): 778.

AtMGT9基因T-DNA插入突变体表型分析

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