非肌肉肌球蛋白Ⅱ型重链A羧基端蛋白的真核表达及在猪繁殖与呼吸综合征病毒感染Marc-145细胞中的作用

Abstract

Abstract: Our previous studies demonstrated that anti-idiotypic antibody against porcine reproductive and respiratory syndrome virus (PRRSV) GP5 protein specifically binds nonmuscle myosin heavy chain II-A (NMHC II-A) on Marc-145 cells and the binding site is in the NMHC II-A C-terminus region (named PRA). In this study, PRA protein was expressed in eukaryotic system and examined for its roles in PRRSV infection of Marc-145 cells. PRA protein containing 310 amino acids was expressed in Bac-to-Bac baculovirus expression system and identified by Western blot. Its binding to Marc-145 cells was detected by indirect immunofluorescence assay. The ability of PRA protein to block PRRSV infection of Marc-145 cells was examined by virus neutralization test, fluorescent focus neutralization test and quantitative RT-PCR. PRA protein was expressed in eukaryotic system with the highest expression level at MOI 1 after 96 hours of infection. The results showed that PRA protein specifically bound with Marc-145 cells and inhibited the PRRSV infectivity up to 60%. Our results indicated that NMHC Ⅱ-A C-terminus protein was successfully expressed in baculovirus expression system and could inhibite the PRRSV infectivity of Marc-145 cells up to 60%. These results provided additional information on the function of NMHC II-A during PRRSV infection.

CLC Number:

S852.659.6

吕Jun-Hua,LIU Ning-Ning, ZHAO Qin, MA Yu-Ping, ZHANG Chong, WANG Xiang-Peng, . Eukaryotic Expression of Nonmuscle Myosin Heavy Chain II-A C-terminus Protein
and Its Roles in Porcine Reproductive and Respiratory Syndrome Virus Infection of
Marc-145 Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2012, 43(9): 1455-1462.

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Eukaryotic Expression of Nonmuscle Myosin Heavy Chain II-A C-terminus Protein
and Its Roles in Porcine Reproductive and Respiratory Syndrome Virus Infection of
Marc-145 Cells

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Eukaryotic Expression of Nonmuscle Myosin Heavy Chain II-A C-terminus Protein and Its Roles in Porcine Reproductive and Respiratory Syndrome Virus Infection of Marc-145 Cells

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Eukaryotic Expression of Nonmuscle Myosin Heavy Chain II-A C-terminus Protein
and Its Roles in Porcine Reproductive and Respiratory Syndrome Virus Infection of
Marc-145 Cells