The upstream regulatory region of a seed specific gene was isolated from the genomic DNA of Brassica napus by PCR amplification. The cloned fragment contained 1755 nucleotides, and shared a sequence homology of 99.6%...The upstream regulatory region of a seed specific gene was isolated from the genomic DNA of Brassica napus by PCR amplification. The cloned fragment contained 1755 nucleotides, and shared a sequence homology of 99.6% with the reported data. The coding region of oleic acid desaturase gene was then cloned from Arabidopsis thaliana. The sequencing analysis indicated that the sequence of the PCR product was just the same as reported before. In addition, the plant expression vector harboring the seed specific promoter and trans Fad2 gene was constructed.展开更多
启动子是基因表达的重要顺式调控元件,在基因工程中,种子特异性启动子可以调控外源基因在种子中特异表达,提高表达效率,增强转基因的效果。本试验根据棉花LEA蛋白D34基因序列设计引物,以海岛棉基因组DNA为模板,通过touch down PCR技术,...启动子是基因表达的重要顺式调控元件,在基因工程中,种子特异性启动子可以调控外源基因在种子中特异表达,提高表达效率,增强转基因的效果。本试验根据棉花LEA蛋白D34基因序列设计引物,以海岛棉基因组DNA为模板,通过touch down PCR技术,获得D34基因的种子特异性启动子片段。测序结果表明该片段长1 384 bp,与已发表的D34基因序列相似度为94.87%。该片段除了含有启动子的基本元件TATA框、CAAT框外,还含有种子特异性启动子元件E-box、G-box、B-box、AACA基序等。此外,对其顺式元件做了生物学功能分析。展开更多
The promoter region (BCSP666) of b-conglycinin a-subunit gene from the genomic DNA of soybean Jilin 43 was isolatedby PCR method. Sequencing analysis showed that the cloned fragment BCSP666 had the similar structure ...The promoter region (BCSP666) of b-conglycinin a-subunit gene from the genomic DNA of soybean Jilin 43 was isolatedby PCR method. Sequencing analysis showed that the cloned fragment BCSP666 had the similar structure to the soybeanseed-specific promoter b-conglycinin a'-subunit gene promoter and b-conglycinin b-subunit gene promoter, and it alsocontains many motifs that contribute to the seed-specific promoter activity. Based on this sequencing analysis, wededuced that promoter fragment BCSP666 had the seed-sepecific promoter activity. And then we constructed the seed-specific expression vector pBMI666 with the promoter fragment BCSP666 and D6-fatty acid desaturase gene fromMortierella isabellina. The D6-fatty acid desaturase is the rate-limiting enzyme of the desaturation of linoleic acid in theproduction of a human essential fatty acid, g-linolenic acid(GLA). The production of g-linolenic acid(GLA) was observedin soybean callus cells, which were transformed with this vector. This confirmed the activity of the activity fragmentBCSP666.展开更多
文摘The upstream regulatory region of a seed specific gene was isolated from the genomic DNA of Brassica napus by PCR amplification. The cloned fragment contained 1755 nucleotides, and shared a sequence homology of 99.6% with the reported data. The coding region of oleic acid desaturase gene was then cloned from Arabidopsis thaliana. The sequencing analysis indicated that the sequence of the PCR product was just the same as reported before. In addition, the plant expression vector harboring the seed specific promoter and trans Fad2 gene was constructed.
文摘启动子是基因表达的重要顺式调控元件,在基因工程中,种子特异性启动子可以调控外源基因在种子中特异表达,提高表达效率,增强转基因的效果。本试验根据棉花LEA蛋白D34基因序列设计引物,以海岛棉基因组DNA为模板,通过touch down PCR技术,获得D34基因的种子特异性启动子片段。测序结果表明该片段长1 384 bp,与已发表的D34基因序列相似度为94.87%。该片段除了含有启动子的基本元件TATA框、CAAT框外,还含有种子特异性启动子元件E-box、G-box、B-box、AACA基序等。此外,对其顺式元件做了生物学功能分析。
基金supported by the National Natural Science Foundation of China(30200176)the Key Foundation of Tianjin of China(013802511)
文摘The promoter region (BCSP666) of b-conglycinin a-subunit gene from the genomic DNA of soybean Jilin 43 was isolatedby PCR method. Sequencing analysis showed that the cloned fragment BCSP666 had the similar structure to the soybeanseed-specific promoter b-conglycinin a'-subunit gene promoter and b-conglycinin b-subunit gene promoter, and it alsocontains many motifs that contribute to the seed-specific promoter activity. Based on this sequencing analysis, wededuced that promoter fragment BCSP666 had the seed-sepecific promoter activity. And then we constructed the seed-specific expression vector pBMI666 with the promoter fragment BCSP666 and D6-fatty acid desaturase gene fromMortierella isabellina. The D6-fatty acid desaturase is the rate-limiting enzyme of the desaturation of linoleic acid in theproduction of a human essential fatty acid, g-linolenic acid(GLA). The production of g-linolenic acid(GLA) was observedin soybean callus cells, which were transformed with this vector. This confirmed the activity of the activity fragmentBCSP666.
