[1]唐玉林,李小杰,钟昀廷,等.利用酵母表达体系研究大豆SALI3-2蛋白功能[J].深圳大学学报理工版,2007,24(3):324-330.
 TANG Yu-lin,LI Xiao-jie,ZHONG Yun-ting,et al.Functional analysis of soybean SALI3-2 in yeast[J].Journal of Shenzhen University Science and Engineering,2007,24(3):324-330.
点击复制

利用酵母表达体系研究大豆SALI3-2蛋白功能()
分享到:

《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第24卷
期数:
2007年3期
页码:
324-330
栏目:
生命科学
出版日期:
2007-07-30

文章信息/Info

Title:
Functional analysis of soybean SALI3-2 in yeast
文章编号:
1000-2618(2007)03-0324-07
作者:
唐玉林李小杰钟昀廷郑易之
深圳大学生命科学学院,深圳市微生物基因工程重点实验室,深圳518060
Author(s):
TANG Yu-linLI Xiao-jieZHONG Yun-tingand ZHENG Yi-zhi
College of Life Sciences,Shenzhen Key Laboratory of Microbial Gene Engineering,Shenzhen University,Shenzhen 518060,P. R. China
关键词:
SALI3-2BURP结构域盐胁迫渗透胁迫信号肽大豆蛋白酵母体系蛋白质表达
Keywords:
SALI3-2BURP-domainsalt stressosmotic stresssignal peptidesoybean proteinyeast systemprotein expression
分类号:
Q 943.2;Q 786
文献标志码:
A
摘要:
以含有大豆Sali3-2 cDNA的重组载体为模版,扩增出SALI3-2蛋白及其缺失信号肽的SI cDNA片段,构建酵母表达载体pYES2/S、pYES2/SI以及可表达SALI3-2-His6的pYES2/SH.空载体pYES2/ct和重组载体在转化酿酒酵母INVSC1后得到相应的酵母重组子INV/CK(对照菌)、INV/S、INV/SI和INV/SH.Western blot实验结果表明,SALI3-2蛋白在酵母INVSC1中可被半乳糖诱导表达.对INV/CK、转基因的重组菌INV/S、INV/SI在无胁迫、含1.5mol/L NaCl或山梨醇培养基中的生长情况进行分析.结果表明,大豆SALI3-2蛋白及其缺失多肽SI的表达对无胁迫条件下酵母的生长没有影响,而在NaCl胁迫下可明显提高酵母转化子的耐盐能力.信号肽的缺失使SALI3-2蛋白的耐盐能力有所下降,可见信号肽对该蛋白耐盐功能的发挥有重要作用.另外,SALI3-2蛋白及其缺失多肽SI的表达不能提高酵母重组子的抗渗透能力,表明SALI3-2蛋白对细胞不具有明显的渗透胁迫调节作用.
Abstract:
Soybean SALI3-2 protein is a member of the BURP-domain family. In this study,the full-length cDNA of SALI3-2 protein and its signal sequence-deleted fragment were cloned into the yeast expression plasmid pYES2/ct to construct the pYES2/S for the expression of SALI3-2,the pYES2/SH for the expression of SALI3-2-His6,and the pYES2/SI for the expression of the C-terminal of SALI3-2 without signal peptide. These constructed vectors as well as the control vector pYES2/ct were respectively transformed into yeast INVSC1 to create recombinants INV/S,INV/SH,INV/SI and the control INV/CK. Western blot analysis revealed that the expression of SALI3-2 could be induced by galactose in the recombinants harboring Sali3-2. Growth curves of all yeast recombinant were analyzed in condition of non-stress,high salinity (1.5 mol/L NaCl) and osmotic (2.2 mol/L sorbitol) stress. Results showed that the expression of SALI3-2 or its deleted peptide SI did not influence the yeast growth in the condition without any stress but could elevate the salt tolerance of yeast under 1.5 mol/L NaCl stress. Signal peptide deletion weakened the salt-tolerant function of SALI3-2. It indicats that the signal peptide is important for the function of this protein. The expression of SALI3-2 protein or its deleted peptide SI could not enhance the osmotic stress-tolerance of yeast transformants. It suggests that SALI3-2 protein can not help cells to counteract osmotic stress.

参考文献/References:

[1]Hattori J,Boutilier K A. 以一保守的BURP结构域定义一类具特殊一级结构的植物新蛋白[J] .分子基因遗传学, 1998,259:424-428 (英文版).
[2]Chang-Hyo Goh,Hong Gil Nam,Yu Shin Park. 植物胁迫记忆:ABA处理的拟南芥经光刺激引起的气孔反应的负调节作用及rd22基因的瞬时响应[J].植物杂志,2003,36:240-255 (英文版).
[3]Zheng L,Heupel RC,DellaPenna D. 番茄果实多聚半乳糖醛酸酶异构酶1的β亚基:分离、特性和独特结构特征分析[J]. 植物细胞,1992, 4:1147-1156 (英文版).
[4]Anthea k,Batchelor,Kim Boutilier,等. SCB1,一个大豆种皮发育相关的 BURP-domain蛋白基因[J]. 植物,2002,215:523-532 (英文版) .
[5]Aiming Wang,Qun Xia,Wen Shuang Xie,等. 典型的乌氏体携带一个以孢子体形式产生的花粉发育所必需的结构蛋白(RAFTIN)[J]. 美国科学院院报,2003, 24:14487-14492 (英文版).
[6]Yu S,Zhang L,Zuo K,等. 从油菜中分离出一个与生物和非生物胁迫有关的BURP domain基因BnBDC1及其特点[J]. 植物生理学,2004, 122:210-218 (英文版).
[7]Ragland M,Soliman KM.? Sali5-4a和 Sali3-2,大豆根部两个铝诱导基因[J]. 植物生理,1997, 114:395 (英文版).
[8]Toshiaki Banzai,Kikuko Sumiya,Nobutaka Hanagta,等. 从红树中克隆到的一编码BURP domain 蛋白的基因及其性质[J]. 树,2002,16:87-93 (英文版).
[9]Fernandez L,Torregrosa L,Terrier N,等 .葡萄果实发育过程中与果肉形态建成有关的基因的鉴定[J]. 植物分子生物学,2007, 63:307-323 (英文版).
[10]Granger C,Coryell V,Khanna A,等. 大豆中多种BURP-domain 蛋白组成员的鉴定、结构及特异性表达[J]. 基因组,2002,45:693-701 (英文版).
[11]余玉雯,孙海丹,郑易之,等. 大豆耐盐相关基因的分离及其功能鉴定[J]. 深圳大学学报理工版,2004,21:324-330.
[12]Lei Zhang,Akinori Ohta,Masamichi Takagi,等.? 植物lea2和lea3基因在酵母中的表达揭示了LEA蛋白的功能多样性[J]. 生物化学杂志,2000,127:611-616 (英文版).
[13]俞嘉宁,张林生,张劲松,等. 小麦耐逆基因-TaLEA3的克隆及在酵母中的功能分析[J]. 生物工程学报, 2004,20(6):832-838.
[14]刘大丽,张欣欣,程玉祥,等. 逆境下水稻(Oryza sativaL.) rHsp90基因的克隆及功能分析[J]. 分子植物育种,2006,4(3):317-322.
[15]Matteo A Di,Federici L,Salvi G,等. 多聚半乳糖醛酸酶抑制蛋白,一个参与植物防御反应的亮氨酸丰富重复蛋白的晶体结构[J].美国科学院院报,2003, 17:10124-10128 (英文版).
[16]Zhu J K. 盐和干旱胁迫信号在植物中的转导[J]. 植物生物学年鉴, 2002,53:247-273(英文版).
[17]叶展辉,郑易之,刘 昀. 大豆PM2蛋白11氨基酸结构域的耐盐功能鉴定[J]. 深圳大学学报理工版,2006,23(4):362-367.
[1]Hattori J,Boutilier K A. A conserved BURP domain defines a novel group of plant proteins with unusual primary structures[J]. Mol Gen Genet,1998,259:424-428.
[2]Chang-Hyo Goh,Hong Gil Nam,Yu Shin Park. Sress memory in plants:a negative regulation of stomatal response and transient induction of rd22 gene to light in abscisic acid-entrained Arabidopsis plants[J]. The Plant Journal,2003,36:240-255.
[3]Zheng L,Heupel RC,DellaPenna D. The bata subunit of tomato fruit polygalacturonase isoenzyme 1:isolation,characterization,and identification of unique structural features[J]. Plant Cell,1992,4:1147-1156.
[4]Anthea k,Batchelor,Kim Boutilier,et al. SCB1,a BURP-domain protein gene,from developing soybean seed coats[J]. Planta,2002,215:523-532.
[5]Aiming Wang,Qun Xia,Wen Shuang Xie,et al. The classical Ubisch bodies carry a sporophytically produced structural protein (RAFTIN) that is essential for pollen development[J]. PANS,2003,24:14487-14492.
[6]Yu S,Zhang L,Zuo K,et al. Isolation and characterization of a BURP domain-containing gene BnBDC1 from Brassica napus involved in abiotic and biotic stress[J]. Physiol Plantarum,2004,122:210-218.
[7]Ragland M,Soliman KM. Sali5-4a and Sali3-2,two genes induced by aluminum in soybean roots[J]. Plant Physiol,1997,114:395.
[8]Toshiaki Banzai,Kikuko Sumiya,Nobutaka Hanagta,et al. Molecular cloning and characterization of genes encoding BURP domain-containing protein in the mangrove,Bruguiera gymmorrhiza[J]. Trees,2002,16:87-93.
[9]Fernandez L,Torregrosa L,Terrier N,et al. Identification of genes associated with flesh morphogenesis during grapevine fruit development[J]. Plant Mol Biol,2007,63:307-323.
[10]Granger C,Coryell V,Khanna A, et al. Identification,structure,and differential expression of members of a BURP domain containing protein family in soybean[J]. Genome,2002,45:693-701.
[11]YU Yu-wen,SUN Hai-dan,ZHENG Yi-zhi,et al. Isolation and characterization of genes related to salt-tolerance in soybean[J]. Journal of Shenzhen University Science and Engineering,2004,21(4):324-330 (in Chinese).
[12]Lei Zhang,Akinori Ohta,Masamichi Takagi,et al. Expression of plant group 2 and group 3 lea genes in Saccharomyces cerevisiae revealed functional divergence among LEA proteins[J]. J Biochem,2000,127:611-616.
[13]YU Jia-ning,ZHANG Lin-sheng,ZhANG Jin-song, et al. Cloning of a novel stress-tolerant gene-TaLEA3 from wheat and the functional analysis in yeast[J]. Chinese Journal of Biotechnology,2004,20(6):832-838 (in Chinese).
[14]LIU Da-li,ZHANG Xin-xin,CHENG Yu-xiang,et al. Cloning and characterization of the rHsp90 gene in rice under environment stress[J]. Molecular Plant Breeding,2006,4(3):317-322 (in Chinese).
[15]Matteo A Di,Federici L,Salvi G,et al. The crystal structure of polygalacturonase-inhibiting protein (PGIP),a leucine-rich repeat protein involved in plant defense[J]. PANS,2003,17:10124-10128.
[16]Zhu JK. Salt and drought stress signal transduction in plants[J]. Annu Rev Plant Biol,2002,53:247-273.
[17]YE Zhan-hui,ZHENG Yi-zhi,LIU Yun. 11-mer repeating region in soybean PM2 protein enhances salt tolerance of Escherichia coli[J]. Journal of Shenzhen University Science and Engineering,2006,23(4):362-367 (in Chinese).

更新日期/Last Update: 2007-12-07