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Purpose expressed in Pichia pastoris GS115 the antimicrobial peptides ABPj-1, detected antibacterial, anti-viral activity. Observed ABPj-1 experimental animal nervous system, the cardiovascular system and the respiratory system; the Acute toxicity observed in experimental animals ABPj-1 long-term toxicity and teratogenicity. Provide experimental basis for clinical. The synthetic ABPj-1 gene sequence and cloned into Pichia pastoris expression vector pPIC9K. Recombinant expression the plasmid pPIC9K-ABPj-1 was transformed into Pichia pastoris GS115 by G418, induced the expression of positive clone strains, the identification of the product and the thermal stability test, challenge test. Detect its antibacterial and antiviral activity. Observed on locomotor activity in mice after oral administration, in order to understand its impact on the nervous system of laboratory animals; observed ABPj-1 in anesthetized dogs blood pressure, heart rate, respiratory rate, and breathing rate and a series of indicators to understand its heart vascular and respiratory systems. Application acute toxicity test, the long-term toxicity tests, systemic allergy test, causing mutation assay and micronucleus test its toxic effects. The results successfully established a recombinant plasmid pPIC9K-ABP-j1, and successful induction of expression in Pichia pastoris GS115, and to the target protein identified by SDS-PAGE. After testing, ABP-j1 has strong thermal stability, and has strong antibacterial and antiviral activity. In this experiment, a single oral gavage ABP-j1 were 2.5mg/kg, 10mg/kg, 40mg/kg normal spontaneous activity did not show excitement or inhibition of state, and sedation with sodium pentobarbital hypnotic no obvious synergy. The anesthetized dogs intravenous injection of 2.5mg/kg, 10mg/kg, 40mg/kg cardiovascular system of laboratory animals and respiratory system had no significant effect. Mice administered acute toxicity test, the test mice without a death, there is no measure LD50 is a non-toxic drug that ABP-j1. Systemic allergy test, the the ABP-j1 Group score of 0, the ABP-j1 allergic. Detected by the micronucleus test and sperm abnormality test, the test results were negative, ABP-j1 not caused by a micro-nuclear role and induced sperm abnormality role. Rats with long-term medication, ABP-j1 low, medium and high dose group were no deaths, generally in good condition, hematologic indicators, blood biochemical indicators and vital organs morphology were not significantly affected, suggesting that ABP-j1 The long-term medication is safe.
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