Improved protein quality in transgenic soybean expressing a de novo synthetic protein, MB-16

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• 作者：Yunfang Zhang (1) (4)
  Johann Schernthaner (1)
  Natalie Labbé (1)
  Mary A. Hefford (2)
  Jiping Zhao (3)
  Daina H. Simmonds (1)
  
• 关键词：Transgenic soybean ; Milk bundle protein ; MB ; 16 ; β ; Conglycinin ; Cruciferin ; Sulfur amino acids ; Essential amino acids
• 刊名：Transgenic Research
• 出版年：2014
• 出版时间：June 2014
• 年：2014
• 卷：23
• 期：3
• 页码：455-467
• 全文大小：
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• 作者单位：Yunfang Zhang (1) (4)
  Johann Schernthaner (1)
  Natalie Labbé (1)
  Mary A. Hefford (2)
  Jiping Zhao (3)
  Daina H. Simmonds (1)
  
  1. Eastern Cereals and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ottawa, ON, K1A0C6, Canada
  4. Shanghai Institute of Biological Science, China Academic Science, 320 Yueyang Road, Life Science Building, Shanghai, 200032, China
  2. Centre for Biologics Research, Biologics and Genetic Therapies Directorate, Health Canada, Ottawa, ON, K1A 0L2, Canada
  3. Ball Helix, Ball Horticultural Company, 622 Town Road, West Chicago, IL, 60185-2698, USA
  
• ISSN：1573-9368

文摘

To improve soybean [Glycine max (L.) Merrill] seed nutritional quality, a synthetic gene, MB-16 was introduced into the soybean genome to boost seed methionine content. MB-16, an 11?kDa de novo protein enriched in the essential amino acids (EAAs) methionine, threonine, lysine and leucine, was originally developed for expression in rumen bacteria. For efficient seed expression, constructs were designed using the soybean codon bias, with and without the KDEL ER retention sequence, and β-conglycinin or cruciferin seed specific protein storage promoters. Homozygous lines, with single locus integrations, were identified for several transgenic events. Transgene transmission and MB-16 protein expression were confirmed to the T5 and T7 generations, respectively. Quantitative RT-PCR analysis of developing seed showed that the transcript peaked in growing seed, 5-?mm long, remained at this peak level to the full-sized green seed and then was significantly reduced in maturing yellow seed. Transformed events carrying constructs with the rumen bacteria codon preference showed the same transcription pattern as those with the soybean codon preference, but the transcript levels were lower at each developmental stage. MB-16 protein levels, as determined by immunoblots, were highest in full-sized green seed but the protein virtually disappeared in mature seed. However, amino acid analysis of mature seed, in the best transgenic line, showed a significant increase of 16.2 and 65.9?% in methionine and cysteine, respectively, as compared to the parent. This indicates that MB-16 elevated the sulfur amino acids, improved the EAA seed profile and confirms that a de novo synthetic gene can enhance the nutritional quality of soybean.