Comparison of expression of monomeric and multimeric adenoregulin genes in Escherichia coli and Pichia pastorias.
Adenoregulin is a 33 amino acid antibiotic peptide who belongs to dermaseptin family which is the first vertebrate family to show lethal effects against filamentous fungi, as well as a broad spectrum of pathogenic microorganisms. Synthetic adenoregulin gene was cloned in 2, 4 and 6 tandem repeats and subcloned in pET32a and pET22b vectors. Recombinant plasmids were transformed into E. coli BL21(DE3), Fusion proteins of Trx-ADR1, Trx-ADR2 and Trx-ADR4 could be expressed after the hosts were induced by IPTG, but the expression level decreased dramatically with the number of tandem repeats increased. ADR1, ADR4 and ADR6 could not be expressed by E. coli without carrier proteins. But for Pichia pastoris GS115, ADR1 and ADR6 in the fermentation broth of the hosts could be detected by ELISA, and the bactericidal activities could also be observed.
Zhou
Yuxun
Y
State Key Laboratory of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, P.R.
Cao
Wei
W
Wang
Jinzhi
J
Ma
Yushu
Y
Wei
Dongzhi
D
D029845:Amphibian Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D000906:Antibodies; D023181:Antimicrobial Cationic Peptides / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005285:Fermentation; D015870:Gene Expression; D010843:Pichia / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D020080:Tandem Repeat Sequences
D029845:Amphibian Proteins; D000906:Antibodies; D023181:Antimicrobial Cationic Peptides; D011994:Recombinant Proteins; C485283:dermaseptin B2, Phyllomedusa bicolor