Isolation and Structural Analysis of the Seed-Specific Promoter from Soybean

来源 :Agricultural Sciences in China | 被引量 : 0次 | 上传用户:liuhaiyi123
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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. The promoter region (BCSP666) of b-conglycinin a-subunit gene from the genomic DNA of soybean Jilin 43 was isolated by 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 from Mortierella isabellina. The D6-fatty acid desaturase is the rate-limiting enzyme of the desaturation of linoleic acid in the production of a human essential fatty acid , g-linolenic acid (GLA). The production of g-linolenic acid (GLA) was observed in soybean callus cells, which were transformed with this vector. This con firmed the activity of the activity fragmentBCSP666.
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