种子特异启动子的克隆及植物表达载体构建Cloning of Seed-specific Promoter and Construction of Plant Transformation Vector
张梦晗;谢伟红;曾洪斌;徐超;李宏业;杨维东;刘洁生;
摘要(Abstract):
[目的]启动子是植物基因工程中一个重要工具,对构建植物生物反应器有着重要意义。8S球蛋白是绿豆中含量最丰富的种子贮藏蛋白,因此,其调控序列可能提供了一个很好来源的种子特异性启动子。[方法]通过基因组步移技术克隆了绿豆8S球蛋白a亚基基因8SGα的启动子区域,基因组步移使用的引物根据绿豆8SGα的cDNA序列设计。[结果]通过3轮PCR的扩增,得到了472 bp的上游序列片段,构建了植物双元表达载体pBI-8SGα-GUS。[结论]研究结果可用于转化植物,并于转基因植物中分析启动表达的时空特异性及表达强度,并为植物种子生物反应器提供重要元件。
关键词(KeyWords): 启动子;PCR;基因组步移;绿豆8S球蛋白;种子特异表达载体
基金项目(Foundation): 广东省自然科学基金面上项目(9151063101000001);; 国家大学生创新性实验计划项目(081055908)
作者(Author): 张梦晗;谢伟红;曾洪斌;徐超;李宏业;杨维东;刘洁生;
Email:
DOI: 10.13989/j.cnki.0517-6611.2010.01.046
参考文献(References):
- [1]STOGER E,VAQUERO C,TORRES E,et al.Cereal crops as viable pro-duction and storage systems for pharmaceutical scFv antibodies[J].PlantMol B iol,2000,42:583-590.
- [2]LI H Y,JIANG L.Seeds as bioreactors for producing biopharmaceuticals[M]//JAIME A,TEIXEIRA DA SILVA.Floriculture,ornamental andplant biotechnology:advances and topical issues(1st ed).UK:Global Sci-ence Books Ltd,2006:39-45.
- [3]YOSHIHARA Y,WASHIDAH,TAKAIWA F.Assessment of common regula-tory regions required for the endosperm specific expression of rice storageprotein glutelin genes by hybrid promoters[J].Plant Sci,1996,121:63-73.
- [4]XUE G P,PATELM,JOHNSON J S,et al.Selectablemarker-free transgen-ic barley producing a high level of cellulase(1,4β--glucanase)in develo-ping grains[J].Plant Cell Rep,2003,21:1088-1094.
- [5]DE JAEGER G,SCHEFER S,JACOBS A,et al.Boosting heterologous pro-tein production in transgenic dicotyledonous seeds using Phaseolus vulgarisregulatory sequences[J].Nat B iotechnol,2002,20(12):1265-1268.
- [6]BEMARDO A E,GARCIA R N,ADACHIM,et al.8S globulin of mung-bean[Vigna radiata(L.)W ilczek]:cloning and characterization of itscDNA isoforms,expression inEscherichia col,i purification,and crystalliza-tion of themajor recombinant 8S isoform[J].JAgric Food Chem,2004,52(9):2552-2560.