Wang Yongbin Guo Changhong Guo Donglin Cai Qiuyi Zhang Shuang Li Jilin Lin Hong.CLONINGOF SOYBEAN FERRITIN GENE AND CONSTRUCTIONOF ITS PLANT EXPRESSION VECTOR[J].Soybean Science,2006,25(04):454-457.[doi:10.11861/j.issn.1000-9841.2006.04.0454]
大豆铁蛋白 cDNA 的克隆及植物表达载体的构建*
- Title:
- CLONINGOF SOYBEAN FERRITIN GENE AND CONSTRUCTIONOF ITS PLANT EXPRESSION VECTOR
- Keywords:
- soybean ferritin ; cDNA ; cloning ; plant expression vector
- 文献标志码:
- A
- 摘要:
- 以大豆为材料 ,利用 RT - PCR法扩增得到大小约 750bp 的片段 ,将这个片段连接到克隆载体 pBlueskript SK +上进行测序 ,结果证明 ,此片段就是大豆的铁蛋白基因。将此片段再正向插入到植物表达载体 pBI121 的 CaMV35s 启动子和 NOS 终止子之间, 构建了植物表达载体 pBIFe ,并成功地将该载体导入根癌农杆菌 EHA105。此项工作为获得表达铁蛋白的转基因植物奠定了基础。
- Abstract:
- A 750bp fragment was amplified by RT-PCR from soybean. The PCR products were cloned intopBlueskript SK +vector and named pFer after its sequences was confirmed. The soybean ferritin gene wassubcloned into plant expression vector pBI121 and named PBI121Fer. Then the recombination plasmidwere introduced into Agrobacterium EHA105. This work provides a foundation for transferring ferritingene into plant by Agrobacterium mediated transformation.
参考文献/References:
1 Andrews S C , Arosio P , Bottke W , et al. Structure, functionand evolution of ferritins [ J] . J Inorg Chem 1992, 47:161 -174.2 Harrison P M , A rosio P. The ferritins:Molecular properties, i-ron storage function and cellular regulation[ J] . Biochim BiophysActa. 1996, 1275:161- 203.3 Goto F , Yoshihara T , Masuda T , T akaiwa F. Genetic improve-ment of iron content and stress adaptation in plants using ferritingene [ J] . Biotechnol Genet Eng Rev. 2001, 18:351- 71.4 Lobreaux S, Massenet O , Briat J F. Iron induces ferritin syhth-sis in maize plantlets [ J] . Plant Mol Biol. 1992, 19(4):563 -5755 Van Wuytswinkel, O, Savino G , Briat J F. Purification andcharacterization of recombinant pea-seed ferritins expressed inEscherichia coli:influence of N-terminus deletions on protein sol-ubility and core formation in vitro[ J] . Biochem. J. 1995, 305:253 - 2616 萨姆布鲁克. 分子克隆实验指南 第三版[ M] 科学出版社, 2002.7 World health Organization http:/ /www. who. int /nut /malnutri-tion-worldwide. htm#ida8 Stearman R, Yuan D S, Yamaguchi-Ivai, Y et al. A permease-oxidase comples involved in high-affinity iron uptake in yeast.Science [ J] . 1996, 271:1552- 1557.9 Gregorio G B. Progress in breeding for trace minerals in staplecrops[ J] . J Nutr, 2002, 132:500 - 502.
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备注/Memo
基金项目:黑龙江自然科学基金 C2005 - 07;哈尔滨市学科后备带头人项目 2005A FXXJ016;黑龙江省青年基金 QC05C60;黑龙江省教育厅海外学人科研项目 1055HQ024