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 LI Yan,WANG Haiyan,ZHANG Yizheng.Localization of IbSUT1x Protein from Ipomoea batatas (L.) Lam in Yeast Cells[J].Chinese Journal of Applied & Environmental Biology,2010,16(06):798-802.[doi:10.3724/SP.J.1145.2010.00798]





Localization of IbSUT1x Protein from Ipomoea batatas (L.) Lam in Yeast Cells
李岩王海燕 张义正
(1四川大学生命科学学院,生物资源与生态环境教育部重点实验室,四川省分子生物学与生物技术重点实验室 成都 610064)
(2山东农业大学农学院,作物生物学国家重点实验室,山东省作物生物学重点实验室 泰安 271018)
LI Yan WANG Haiyan ZHANG Yizheng
(1Key Laboratory of Biology Resource & Eco-environment, Ministry of Education, Sichuan Key Laboratory of Molecular Biology & Biotechnology,
College of Life Sciences, Sichuan University, Chengdu 610064, China)
(2State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Taian 271018, China)
Ipomoea batatas sucrose transporter gene expression localization GFP Saccharomyces cerevisiae
蔗糖转运蛋白是植物特有的一类载体蛋白,酿酒酵母突变株SUSY7/ura3常被利用来研究蔗糖转运蛋白的功能,但在研究中发现,有的蔗糖转运蛋白在酵母中未表现出蔗糖转运的功能. 但这或许并不是其实际功能的体现,而是由于这类蛋白无法准确定位到酵母的细胞膜上造成的. IbSUT1x是本实验室从甘薯中分离到的一个蔗糖转运蛋白基因,但是在SUSY7/ura3中未表现出明显的蔗糖转运功能. 本研究通过用激光共聚焦显微镜对IbSUT1x与绿色荧光蛋白(Green fluorescent protein,GFP)的融合蛋白的表达和定位的观察,发现IbSUT1x不仅在酵母细胞中有表达,而且表达的蛋白质定位在酵母的细胞膜上. 结果表明,IbSUT1x在酵母细胞中不能转运蔗糖,该蛋白很可能不具有蔗糖转运功能,或者其与蔗糖的亲和力很低. 图4 表1 参21
Sucrose transporters (SUTs), or termed as sucrose carriers (SUCs), only exist in plants and play important roles in long-distance transport of photosynthate through phloem from source organs to sink ones. Its function was always studied with yeast (Saccharomyces cerevisiae) strain SUSY7/ura3 which is usually used as a model for sucrose uptake activity research. Researchers found that some sucrose transporters could not transport sucrose, which might be on account of the failed localization (not target to plasma membrane) in yeast cells. IbSUT1x is a sucrose transporter gene isolated from sweet potato (Ipomoea batatas), and it does not display apparent sucrose uptake activity. In the present study, it was revealed that IbSUT1x could be successfully targeted to plasma membrane of yeast cells by the localization of IbSUT1x-green fluorescent protein (GFP) fusion protein using laser scanning confocal microscopy. The result suggestes that IbSUT1x can not load sucrose into yeast cells, or just has low sucrose affinity. Fig 4, Tab 1, Ref 21


1 Lalonde S, Wipf D, Frommer WB. Transport mechanisms for organic forms of carbon and nitrogen between source and sink. Annu Rev Plant Biol, 2004, 55: 341~372
2 Sauer N. Molecular physiology of higher plant sucrose transporters. FEBS Lett, 2007, 581: 2309~2317
3 Lemoine R. Sucrose transporters in plants: update on function and structure. Biochim Biophys Acta, 2000, 1465: 246~262
4 Kühn C, Hajirezaei M-R, Fernie AR, Roessner-Tunali U, Czechowski T, Hirner B, Frommer WB. The sucrose transporter StSUT1 localizes to sieve elements in potato tuber phloem and influences tuber physiology and development. Plant Physiol, 2003, 131: 102~113
5 Zhou Y, Qu H, Dibley KE, Offler CE, Patrick JW. A suite of sucrose transporters expressed in coats of developing legume seeds includes novel pH-independent facilitators. Plant J, 2007, 49: 750~764
6 Riesmeier JW, Willmitzer L, Frommer WB. Isolation and characterization of a sucrose carrier cDNA from spinach by functional expression in yeast. EMBO J, 1992, 11 (13): 4705~4713
7 Lemoine R, Bürkle L, Barker L, Sakr S, Kühn C, Regnacq M, Gaillard C, Delrot S, Frommer WB. Identification of a pollen-specific sucrose transporter-like protein NtSUT3 from tobacco. FEBS Lett, 1999, 454: 325~330
8 Li Y, Gu Y, Qin H, Zhang Y. Two pairs of sucrose transporters in Ipomoea batatas (L.) Lam are predominantly expressed in sink leaves and source leaves respectively, Plant Sci, 2010, 179: 250~256
9 Newstead S, Kim H, von Heijne G, Iwata S, Drew D. High-throughput fluorescent-based optimization of eukaryotic membrane protein overexpression and purification in Saccharomyces cerevisiae. Proc Natl Acad Sci USA, 2007, 104 (35): 13936~13941
10 Drew D, Newstead S, Sonoda Y, Kim H, von Heijne G, Iwata GS. GFP-based optimization scheme for the overexpression and puri?cation of eukaryotic membrane proteins in Saccharomyces cerevisiae. Nat Protoc, 2008, 3 (5): 784~798
11 Gietz RD, Schiestl RH. High-ef?ciency yeast transformation using the LiAc/SS carrier DNA/PEG method. Nat Protoc, 2007, 2 (1): 31~34
12 Ausubel FM (奥斯伯), Kingston RE (金斯顿), Scheidemann JG (赛德曼). Short Protocols in Molecular Biology. 4th ed. Beijing, China: Science Press (北京: 科学出版社), 2005
13 Bürkle L, Hibberd JM, Quick WP, Kühn C, Hirner B, Frommer WB. The H+-sucrose cotransporter NtSUT1 is essential for sugar export from tobacco leaves. Plant Physiol, 1998, 118: 59~68
14 Riesmeier JW, Hirner B, Frommer WB. Potato sucrose transporter expression in minor veins indicates a role in phloem loading. Plant Cell, 1993, 5: 1591~1598
15 Shakya R, Sturm A. Characterization of source- and sink-specific sucrose/H+ symporters from carrot. Plant Physiol, 1998, 118: 1473~1480
16 Schmitt B, Stadler R, Sauer N. Immunolocalization of solanaceous SUT1 proteins in companion cells and xylem parenchyma: New perspectives for phloem loading and transport. Plant Physiol, 2008, 148: 187~199
17 Stadler R, Brandner J, Schulz A, Gahrtz M, Sauer N. Phloem loading by the PmSUC2 sucrose carrier from Plantago major occurs into companion cells. Plant Cell, 1995, 7: 1545~1554
18 Stadler R, Sauer N. The Arabidopsis thaliana AtSUC2 gene is specifically expressed in companion cells. Bot Acta, 1996, 109: 299~306
19 Truernit E, Sauer N. The promoter of the Arabidopsis thaliana SUC2 sucrose-H+ symporter gene directs expression of p-glucuronidase to the phloem: Evidence for phloem loading and unloading by SUC2. Planta, 1995, 196: 564~570
20 Schulze W, Weise A, Frommer WB, Ward JM. Function of the cytosolic N-terminus of sucrose transporter AtSUT2 in substrate affinity. FEBS Lett, 2000, 485: 189~194
21 Lu JM-Y, Bush DR. His-65 in the proton-sucrose symporter is an essential amino acid whose modification with site-directed mutagenesis increases transport activity. Proc Natl Acad Sci USA, 1998, 95: 9025~9030


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 TAO Xiang,ZHANG Yongwei,JIANG Yusong,et al.Changes in Root Starch Contents of Sweet Potato Cultivars During Storage[J].Chinese Journal of Applied & Environmental Biology,2010,16(06):741.[doi:10.3724/SP.J.1145.2010.00741]
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国家科技支撑计划项目(No. 2007BAD78B03)和四川省“十一五”重点攻关项目(No. 07SG111-003-1)资助
更新日期/Last Update: 2010-12-29