A novel technique for the effective production of short peptide analogs from concatameric short peptide multimers.
We designed a basic unit of the modified chicken gonadotropin releasing hormone II (cGnRH-II) peptide containing a trypsin cleavable linker peptide at both ends of the original peptide. We made a synthetic DNA coding for the modified cGnRH-II peptide with asymmetric and complementary cohesive ends of linker nucleotides. A tandemly repeated DNA cassette for the expression of concatameric short peptide multimers was constructed by ligating the basic units. The expressed peptide multimers were purified and subject to amino-terminal sequence analysis, which displayed the amino acid sequences expected from the designed nucleotides of the expression cassette. The monomeric cGnRH-II peptide analogs were generated after trypsin digestion. The present results showed that the technique developed for the production of the concatameric peptide multimers with cleavable linker peptides can be generally applicable to the production of short peptide analogs.
Lee
S J
SJ
Biotechnology Division, National Fisheries Research & Development Institute, Pusan, Korea.
Lee
J H
JH
Jin
H J
HJ
Lee
J H
JH
Ryu
H Y
HY
Kim
Y
Y
Kong
I S
IS
Kim
K W
KW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D004274:DNA, Recombinant; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D007987:Gonadotropin-Releasing Hormone / Q000031:analogs & derivatives* / Q000096:biosynthesis* / Q000235:genetics*; D008969:Molecular Sequence Data; D010455:Peptides / Q000138:chemical synthesis* / Q000235:genetics*; D015202:Protein Engineering / Q000379:methods*; D011499:Protein Processing, Post-Translational; D020080:Tandem Repeat Sequences
D004274:DNA, Recombinant; D010455:Peptides; D007987:Gonadotropin-Releasing Hormone; C047717:LHRH, His(5)-Trp(7)-Tyr(8)-