|
KMID : 1007519970060040314
|
|
Food Science and Biotechnology
1997 Volume.6 No. 4 p.314 ~ p.318
|
|
|
Expression and Purification of Soybean Proglycinins Modified by Protein Engineering
|
|
Lee, Sam Pin
Kim, Chan Shick/Ko, Se Kwang/Kang, Soon Sean
|
|
|
Abstract
|
|
|
|
To improve the functional properties of soybean proglycinins, cDNA encoding soybean proglycinin was manipulated using synthetic oligonucleotides. Modified expression plasmid pKGA, _(1a)B_(1b)-3 was used as a template for the insertional mutation. The synthetic oligonucleotides encoding amino acid sequences of Gln-Leu-Ile-Phe-Lys-Leu and Gln-Leu-Phe-Asp-Gln-Thr-Pro-Arg-Val-Phe, respectively, were selectively inserted between Pro^(467) and Gln^(468) in the variable domain V of the clones. The insertion of a new oligonucleotide sequence in recombinant plasmid pKG5S and pKG5P was confirmed by the presence of a PvuII restriction site additionally created. Recombinant plasmids carrying modified cDNA were overexpressed in Escherichia coli JM105. Modified soybean proglycinins are soluble proteins; thus, they were purified by consecutive treatments of salt precipitation, ion-exchange chromatography and cryoprecipitation. Purified proglycinins were confirmed via Western blotting analysis.
|
|
|
KEYWORD
|
|
|
|
|
|
FullTexts / Linksout information
|
|
|
|
|
|
Listed journal information
|
|
 
|
|