To investigate the function of C-terminus nonmuscle myosin heavy chain Ⅱ-A (NMHC Ⅱ -A) to porcine reproductive and respiratory syndrome virus (PRRSV) infection of Marc-145 cells, a part of NMHC II -A gene (PRA) from Marc-145 cells was amplified by RT-PCR and cloned into pEGFP-N1 vector to construct recombinant plasmid pEGFP-N1-PRA for PRA overexpression in Marc-145 cells. The results showed that the repliccation of PRRSV in the pEGFP-N1-PRA transfected Marc-145 cells was decreased by 50% in comparison with non-transfected cells by TCID50 titration and immunofluorescent assay. These results showed PRA encoding protein could influent PRRSV infection to Marc-145 cells. This rinding provide a new way for further understanding of PRRSV infection to permissive cells.%为研究非肌肉肌球蛋白II型重链A(NMHC II -A)羧基端在猪繁殖与呼吸综合征病毒(PRRSV)感染Marc- 145细胞中的作用,本实验通过提取Marc-145细胞总RNA,RT-PCR方法扩增NMHC II -A羧基端930 bp基因PRA,并进行核苷酸序列测定.将该基因连接到pEGFP-N1载体中,采用双酶切和PCR方法对重组质粒pEGFP-N1-PRA进行鉴定.将pEGFP-N1 -PRA转染至Marc-145细胞后感染PRRSV,测定TCID50值及间接免疫荧光(IFA)检测PRA对PRRSV感染Marc- 145细胞的影响.TCID50及IFA结果显示,与未转染的Marc- 145细胞相比,转染pEGFP-N1-PRA的Marc-145细胞的PRRSV感染能力降低50％.本研究初步证明了PRA编码蛋白在PRRSV感染细胞中的作用,为PRRSV感染机制的研究提供了一定的依据.
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