Signal sequence of preproglycinin affects production of the expressed protein in Escherichia coli.
The effect of the signal peptide portion on the bacterial production of preproglycinin, a precursor of soybean storage protein, was examined. Nucleotide sequences corresponding to the signal peptide and the mature N-terminal region were deleted stepwise from the cDNA encoding the glycinin A1aB1b subunit precursor, and the deleted cDNAs were placed under the control of trc promoter in an expression vector pKK233-2. When the amounts of the protein products in Escherichia coli from each expression plasmid were determined, no accumulation of preproglycinin was observed from the plasmids with the full length or the five amino acids of the signal sequence. However, significant accumulation of the preproglycinin homologue proteins was noted from the plasmids retaining less than three amino acids of the signal sequence depending on the extent of deletion. N-terminal amino acid sequences of the products coincided with those predicted from the deleted cDNAs. The preproglycinin homologue proteins expressed from the mutant plasmids assembled into trimers of about 8S.
Utsumi
S
S
Research Institute for Food Science, Kyoto University, Japan.
Kim
C S
CS
Sato
T
T
Kito
M
M
D000595:Amino Acid Sequence; D001483:Base Sequence; D002872:Chromosome Deletion; D003470:Culture Media; D004926:Escherichia coli / Q000235:genetics*; D005786:Gene Expression Regulation; D005916:Globulins / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D011498:Protein Precursors / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics*; D030262:Soybean Proteins
D003470:Culture Media; D005916:Globulins; D011498:Protein Precursors; D021382:Protein Sorting Signals; D030262:Soybean Proteins; C011474:glycinin