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巴西坚果2S白蛋白在甘蓝型油菜中的表达。

Expression of the 2S albumin from Bertholletia excelsa in Brassica napus.

作者信息

Guerche P, De Almeida E R, Schwarztein M A, Gander E, Krebbers E, Pelletier G

机构信息

Laboratoire de Biologie Cellulaire, I.N.R.A., Versailles, France.

出版信息

Mol Gen Genet. 1990 May;221(3):306-14. doi: 10.1007/BF00259393.

Abstract

The methionine rich 2S albumin seed storage protein of Bertholletia excelsa has been expressed in seeds of Brassica napus (rapeseed). A chimeric gene driven by the soybean lectin 5' flanking regions was used to produce a fusion protein consisting of the soybean lectin signal peptide and the propeptide of the Brazil nut 2S albumin. Several transgenic plants were studied at the RNA and protein levels; in each case the chimeric gene was expressed and the protein detected at levels ranging from 0.02% to 0.06% of total protein. Transcriptional studies in a particular transgenic plant show that expression of the gene is tissue specific and developmentally regulated during seed maturation. The endogenous napin genes and the introduced gene are regulated differently, with expression of the chimeric gene paralleling that seen when the soybean lectin gene is expressed in other plant species. Western analysis using antibodies to Brazil nut 2S albumins resulted in the detection of a protein whose size is consistent with correct processing of the precursor.

摘要

巴西坚果的富含甲硫氨酸的2S白蛋白种子贮藏蛋白已在甘蓝型油菜(油菜籽)种子中表达。一个由大豆凝集素5'侧翼区驱动的嵌合基因被用于产生一种融合蛋白,该融合蛋白由大豆凝集素信号肽和巴西坚果2S白蛋白的前肽组成。对多个转基因植株进行了RNA和蛋白质水平的研究;在每种情况下,嵌合基因均表达,且检测到的蛋白质含量占总蛋白的0.02%至0.06%。对一株特定转基因植株的转录研究表明,该基因的表达具有组织特异性,且在种子成熟过程中受发育调控。内源性napin基因和导入基因的调控方式不同,嵌合基因的表达与大豆凝集素基因在其他植物物种中表达时的情况相似。使用针对巴西坚果2S白蛋白的抗体进行的Western分析检测到一种蛋白质,其大小与前体的正确加工一致。

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