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大豆科学  2007 

二维电泳技术在大豆子叶微粒体膜结合蛋白研究中的应用[JP]

DOI: 10.3969/j.issn.1000-9841.2007.01.004, PP. 11-15

Keywords: 二维电泳,大豆,子叶,微粒体膜结合蛋白

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Abstract:

以硝酸银诱导的大豆子叶为材料,初步探讨了二维电泳技术在分离大豆子叶微粒体膜结合蛋白中的应用,研究了重新水化溶液的组成、加样量、等电聚焦的totalvoltagehours以及不同pH梯度范围IPG(固相pH梯度)胶条等因素对膜蛋白的溶解和二维电泳图像的影响。结果表明对于70×3×0.5mmIPG胶条,能够溶解较多蛋白质和获得高质量二维电泳图像的优化条件为:由尿素、硫脲、三丁基膦和C7BZO组成的重新水化溶液(TUC-7T-2)、80μg的加样量和20000vhr的totalvoltagehour。在此优化条件下,与宽范围pH梯度IPG胶条相比,使用窄范围pH梯度胶条可以观察到更多的和质量更好的蛋白质点,结果发现至少有13个点只存在于硝酸银诱导的样品中,而不存在于对照中,其代表的蛋白质的等电点在4.0~8.0。

References

[1]  [1]Welle R., Grisebach H. Properties and solubilization of the prenyltransferase of isoflavonoid phytoalexin biosynthesis in soybean [J]. Phytochemistry, 1991, 30(2):479-484.
[2]  [2]Z.hringer U., Ebel J., Mulheirn L. J.,et al. Induction of phytoalexin synthesis in soybean. Dimethylallylpyrophosphate: trihydroxypterocarpan dimethylallyl transferase from elicitor-induced cotyledons [J]. FEBS letters, 1979, 101(1):90-92.
[3]  [3]Z.hringer U., Schaller E., Grisebach H. Induction of phytoalexin synthesis in soybean. Structure and reaction of naturally occurring and enzymatically prepared prenylated pterocarpans from elicitor-treated cotyledons and cell cultures of soybean [J]. Zeitschrift fuer naturforschung, 1981, 36c, 234-241.
[4]  [4]Ames G. F. L., Nikaido K. Two dimensional gel electrophoresis of membrane proteins [J]. Biochemistry, 1976, 15(3):616-623.[JP].
[5]  [5]Lehner I., Niehof M., Boriak J. An optimized method for the isolation and identification of membrane proteins [J]. Electrophoresis, 2003, 24:1795-1808.
[6]  [6]Santoni V., Molloy M., Rabilloud T. Membrane proteins and proteomics: Un amour impossible [J]. Electrophoresis, 2000, 21:1054-1070.
[7]  [7]乞永艳,Arnaud Bovy, Ric de Vos, 等.外源茉莉酸对大豆中异戊烯黄酮Glyceollins及其前体Glycinol累积的影响[J]大豆科学,2005,24(4):269-273.
[8]  [8]Diesperger H., Müller C. R., Sandermann H. Rapid isolation of a plant microsomal fraction by Mg2+―precipitation [J]. FEBS Letters, 1974, 43(2):155-158.
[9]  [9]Berkelman T., Stenstedt T. 2-D Electrophoresis Principles and Methods [P]Amersham Biosciences, 1998, 80-6429-60 Edition AB.
[10]  [10]Amersham Pharmacia Biotech Immobiline Drystrip short instruction [P]. 71-5009-57 Edition AA, 99-04,1999.
[11]  [11]Amersham Biosciences, Immobiline Drystrip short instruction [P]. 71-5019-33, Edition AD, 03-01,2003.
[12]  [12]Gorg A., Obermaier C., Boguth G., et al. The current state of two dimension electrophoresis with immobilized pH gradients [J]. Electrophoresis, 2000, 21:1037-1053.
[13]  [13]Stossel P. Glyceollin production in soybean [J].Phytopath. Z., 1982, 105:109-119.
[14]  [14]乞永艳.大豆子叶异黄酮类次生代谢物的诱导及其微粒体部分蛋白的初步研究[D]中国农业科学院博士学位论文,2005年12月.

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