Total RNA was extracted from chicken oviduct epithelial cells. Avian Beta-defensin (AvBD)-2, -6, and -12 cDNAs were amplified by reverse transcription-PCR and cloned into pRSET A style='msoareast-language:ZH-CN'>, a protein expression vector. The class=SpellE>hexa-histidine-tagged class=SpellE>AvBD peptides were expressed in Escherichia coli (E. coli) BL21(DE3) class=SpellE>plysS and affinity-purified. The antimicrobial activities of the recombinant AvBDs against E. coli style='msoareast-language:ZH-CN;mso-bidiont-style:italic'>, Salmonella class=SpellE>enterica style='mso-bookmark:OLE_LINK9'> serovar Typhimurium (S. class=SpellE>typhimurium), and Staphylococcus aureus style='msoareast-language:ZH-CN'> (S. aureus) were determined. style='msoareast-language:ZH-CN;mso-bidiont-style:italic'> At 8, 16 and 32 µg/ml, all three rAvBDs killed and inhibited the growth style='msoareast-language:ZH-CN'> of E. coli style='msoareast-language:ZH-CN;mso-bidiont-style:italic'>, S. typhimurium, and S. aureus. The killing of rAvBD-2, -6, and -12 against stationary phase E. coli and S. class=SpellE>aureus was pH dependent in the range investigated. style='msoareast-language:ZH-CN'> In addition, the killing-curves showed that rAvBDs exerted their antimicrobial function within 30 minutes of treatment, suggesting the fast killing mechanisms of rAvBDs.
| Identifer | oai:union.ndltd.org:MSSTATE/oai:scholarsjunction.msstate.edu:td-3189 |
| Date | 30 April 2011 |
| Creators | Zhao, Li |
| Publisher | Scholars Junction |
| Source Sets | Mississippi State University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Theses and Dissertations |
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