金黄色葡萄球菌PV-杀白细胞素的表达及其对奶牛乳腺上皮细胞的损伤作用

doi:10.11843/j.issn.0366-6964.2019.10.017

Abstract

Abstract: In order to provide a basis for systematically studying the mechanism of Staphylococcus aureus infection of bovine mammary epithelial cells, the damages of producing panton-valenine leukocidin (PVL) S. aureus ATCC49775, pvl-knockout mutant △pvl 49775, complemented mutant C-△pvl 49775 and recombinant PVL (rPVL) in vitro on bovine mammary epithelial cells were studied. Recombinant plasmids of S. aureus LukS-PV and LukF-PV were transformed into E. coli BL21, respectively. After expression and purification, they were identified by Western blot. The effects of ATCC49775, △pvl 49775, C-△pvl 49775 and rPVL on the apoptosis of bovine mammary epithelial cells were detected by Live/Dead cell fluorescence staining and Annexin V-FITC/PI double staining. The recombinant expression vectors pET28a-LukS-PV and pET28a-LukF-PV were successfully constructed. The concentration of LukS-PV obtained after induction and purification was 0.6 mg·mL^-1, and the concentration of LukF-PV was 1.15 mg·mL^-1. rPVL mainly induced apoptosis (P<0.01). Both S. aureus ATCC49775, △pvl 49775 and C-△pvl 49775 induced significant apoptosis and necrosis on bovine mammary epithelial cells, but ATCC49775 and C-△pvl 49775 induced worse (P<0.05). The results showed that rPVL of prokaryotic expression had a strong toxicity on bovine mammary epithelial cells. The pvl-knockout mutant △pvl 49775 induced the apoptosis and necrosis of the bovine mammary epithelial cells low than the pvl positive strain ATCC49775 and C-△pvl 49775. Our study indicates that the PVL plays an important role in S. aureus infected bovine mammary epithelial cells.

CLC Number:

S852.611

MA Weiwu, ZHOU Xuezhang. Expression of Panton-Valenine Leukocidin from Staphylococcus aureus and Its Effects on Bovine Mammary Epithelial Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(10): 2105-2112.

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Expression of Panton-Valenine Leukocidin from Staphylococcus aureus and Its Effects on Bovine Mammary Epithelial Cells

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