[Expression of porcine beta-defensin 1 gene in Pichia pastoris].
PBD-1 is an antibacterial peptide that plays an important role in defence system of porcine. To produce PBD-1 with bioactivity in Pichia pastoris, according to published amino acid sequence of porcine beta-defensin 1(PBD-1) and the partiality codon of yeast, the PBD-1 gene was synthesized by PCR and cloned into pPIC9K to construct the recombinant expression vector pPIC9K-PBD-1, the obtained recombinant plasmid was linearized by Sal I, and then transformed into SMD1168 by electroporation. Under the control of the promoter AOX1, an approximately 4.5 kD PBD-1 peptide was expressed. Antibacterial activity assay shows that the PBD-1 has the antibacterial activity on Staphylococcus aureus. This is the first secreted expression of porcine beta-defensin 1 gene in Pichia pastoris.
Jiang
Li-Hua
LH
National Engineering Center for Biotechnology, Shenzhen 518057, China.
Lu
Hai-Rong
HR
Huang
De-Xin
DX
Yi
Jun-Bo
JB
Li
Ling-Yun
LY
Lin
Feng
F
D000818:Animals; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D010843:Pichia / Q000235:genetics*; D015202:Protein Engineering; D013211:Staphylococcus aureus / Q000187:drug effects; D013552:Swine / Q000235:genetics*; D023083:beta-Defensins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology
D000900:Anti-Bacterial Agents; C406407:DEFB1 protein, human; D023083:beta-Defensins