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United States Department of Agriculture

Agricultural Research Service

Title: The Expression of a Synthetic Codon Adjusted Porcine Alpha-Lactalbumin Gene in Maize

item Moran, Daniel
item Scott, Marvin

Submitted to: Maize Genetics Conference Abstracts
Publication Type: Abstract Only
Publication Acceptance Date: March 11, 1999
Publication Date: N/A

Technical Abstract: The main nutritional limitation of corn used in production of swine feed is the content of "ideal protein" that is digestible, bioavailable and contains an amino acid balance that matches the requirements of swine. Porcine alpha-lactalbumin is a major component of sow's milk with high digestibility and excellent amino acid profile with high content of Lys and dTrp. As an initial effort to create corn with a higher content of ideal protein, transgenic maize expressing porcine alpha-lactalbumin have been generated. A codon-adjusted porcine alpha-lactalbumin gene that contains nucleotide sequence for 27 kDa gamma-zein signal peptide at the 5' end was synthesized using recursive PCR method. A plant expression vector that contains codon-adjusted porcine alpha-lactalbumin gene and BAR gene under the cntrol of the maize Ubi- I promoter including first exon and intron and nos3 terminator were constructed. Typell embryogenic callus line was transformed by particle-bombardment-mediated gene transfer. After transformation, transgene integration was verified using PCR among callus lines that were resistant to Bialaphos. A polyclonal antibody against human alpha-lactalbumin raised in rabbits was used for immuno-blot analysis to detect porcine alpha-lactalbumin expression in callus. Transgenic plants were regenerated from callus lines expressing porcine alpha- lactalbumin. Immuno-blot analysis detected porcine alpha-lactalbumin accumulation in whole plant level including leaves, tassel, stems and roots.

Last Modified: 4/22/2015
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