Molecular cloning of an Arabidopsis cDNA encoding a dynamin-like protein that is localized to plastids

被引:51
作者
Kang S.G. [1 ]
Jin J.B. [1 ]
Piao H.L. [1 ]
Pih K.T. [2 ]
Jang H.J. [1 ]
Lim J.H. [1 ]
Hwang I. [1 ,2 ]
机构
[1] Department of Molecular Biology, Gyeongsang National University, Chinju
[2] Plant Molecular Biology and Biotechnology Research Center, Gyeongsang National University, Chinju
关键词
Arabidopsis dynamin-like 2; Chloroplast targeting; GFP fusion protein; Localization;
D O I
10.1023/A:1006099718761
中图分类号
学科分类号
摘要
Dynamin-related proteins are high molecular weight GTPase proteins found in a variety of eukaryotic cells from yeast to human. They are involved in diverse biological processes that include endocytosis in animal cells and vacuolar protein sorting in yeast. We isolated a new gene, ADL2, that encodes a dynamin-like protein in Arabidopsis. The ADL2 cDNA is 2.68 kb in size and has an open reading frame for 809 amino acid residues with a calculated molecular mass of 90 kDa. Sequence analysis of ADL2 revealed a high degree of amino acid sequence similarity to other members of the dynamin superfamily. Among those members ADL2 was most closely related to Dnm1p of yeast and thus appears to be a member of the Vps1p subfamily. Expression studies showed that the ADL2 gene is widely expressed in various tissues with highest expression in flower tissues. In vivo targeting experiments showed that ADL2:smGFP fusion protein is localized to chloroplasts in soybean photoautroph cells. In addition experiments with deletion constructs revealed that the N-terminal 35 amino acid residues were sufficient to direct the smGFP into chloroplasts in tobacco protoplasts when expressed as a fusion protein.
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页码:437 / 447
页数:10
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