Search Keywords: pPICZA Search date: 1/30/06

Interesting Abstracts:

• Wei Sheng Wu Xue Bao. 2005 Apr;45(2):218-22. [Modification of hybrid antimicrobial peptide CecA? -mil gene and its over-secretion expression in Pichia pastoris] [Article in Chinese]

Zhang SF, Cao RB, Jia Y, Zhou B, Chen PY.

Key Laboratory of Animal Disease Diagnosis and Immunology, Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China. zsf780331@tom.com

According to the partiality codon of Pichia pastoris, hybrid antimicrobial peptide CecA? -mil gene was reconstructed, synthesized and cloned into pPICZalpha-A to construct the recombinant expression vector pPICZa-A-CM. The pPICZalpha-A-CM was transformed into yeast host strain X-33. Under the control of the promoter AOX1 (alcohol oxidase 1), a approximately 1.9 kD cecA-mil protein was expressed with the high level of 245 microg/mL after optimized the requirements for the flask-shaking culture fermentation of the Pichia pastoris rX-33/pPICZalpha-A-CM. The hybrid antibacterial peptide had a broad spectrum antibacterial activity on both gram-positive and gram-negative bacteria, especially showed potent antibacterial activity against ampicillin-resistant and kanamycin- resistant bacteria, such as Staphylococcus aureus (cowan I) and pathogenic E. coli (O1) from chicken. In addition, the hybrid antibacterial peptide showed an extreme heat-stable and acid-stable characteristic. Based on these characteristics, the recombinant antibacterial peptide CecA? -mil display application foreground in the field of antisepsis of food, prevention of disease, additives of animal feedstuff and so on.

PMID: 15989264 [PubMed - indexed for MEDLINE]

• Vet Immunol Immunopathol. 2000 Dec 29;77(3-4):233-41. Expression and characterization of a recombinant soluble form of bovine tumor necrosis factor receptor type I.

Mwangi SM, Stabel J, Lee E, Kehrli ME, Taylor MJ.

Biomedical Sciences Department, College of Veterinary Medicine, Iowa State University, Ames, IA 50010, USA.

A recombinant soluble bovine tumor necrosis factor receptor type I (sboTNF-RI) was expressed in the methylotrophic yeast Pichia pastoris and evaluated for its ability to inhibit bovine tumor necrosis factor alpha (TNF-alpha) cytotoxicity. A cDNA encoding the extracellular domain of bovine TNF-RI was placed under the control of the powerful and tightly regulated alcohol oxidase1 (AOX1) gene promoter of the pPICZa A vector and the resulting construct integrated into the 5' region of the alcohol oxidase genes of GS115 and KM71 strains of Pichia. Soluble bovine TNF-RI was secreted into the medium following induction of the AOX1 gene promoter with methanol, and purified to greater than 95% purity by ion-exchange chromatography. In in vitro assays, the purified recombinant sboTNF-RI will block the cytolytic activity of bovine TNF-alpha on WEHI 164 cells clone 13 by 50% when used at a concentration of 170 microg/ml, and by nearly 90% when used at a concentration of 310 microg/ml. Results of this study suggest that recombinant sboTNF-RI may have therapeutic value as a TNF inhibitor in cattle with coliform mastitis.

PMID: 11137122 [PubMed - indexed for MEDLINE]

-- DaniPershouse - 30 Jan 2007