中国兽医科学2018，Vol.48Issue(5)：537-544,8.DOI:10.16656/j.issn.1673-4696.2018.0088
LncRNA TCONS_00179042对Marc-145细胞中PRRSV复制的影响
Effect of LncRNA TCONS_00179042 on the replication of porcine reproductive and respiratory syndrome virus in Marc-145 cells
摘要
Abstract
Marc-145 cells ectopically expressing LncRNA TCONS_00179042 were infected with porcine reproductive and respiratory syndrome virus (PRRSV) GSWW15 strain in order to observe the effect of LncRNA TCONS_00179042 on PRRSV GSWW15 replication. The full-length LncRNA TCONS_00179042 was amplified by RT-PCR. The effect of overexpression of LncRNA TCONS 00179042 on replication of PRRSV GSWW15 strain in the Marc-145 cells was detected by qPCR,TCID50 and Western-blot. The distribution of LncRNA TCONS_00179042 in porcine alveolar macrophages was analyzed by nuclear fraction isolation experiments. The qPCR results demonstrated that the relative expression level of LncRNA TCONS_00179042 in the pcDNA3.1-TCONS_00179042 transfected Marc-145 cells was about 3X 105 folds of the control group,indicating that the LncRNA TCONS_ 00179042 was overexpressed in the Marc-145 cells. The qPCR results also showed that the PRRSV RNA copy number of the overexpressing LncRNA group was about 60％ of that of the control group after infection with the PRRSV GSWW15 strain. In consistent with virus RNA level,the titer of PRRSV in TCONS_00179042 over-expression cells was significantly lower than that of the control cells. The result of Western-blot showed that the expression level of PRRSV N protein in LncRNA over-expression group was significantly lower than that in the control group. These results suggest that LncRNA TCONS_00179042 inhibits the replication of PRRSV GSWW15 strain in Marc-145 cells. The nuclear separation experiment showed that the LncRNA TCONS_00179042 was located both in the cytoplasm and the nucleus, and the expression level in the nucleus was higher than that in the cytoplasm. The overexpression experiments demonstrated that LncRNA TCONS_00179042 has an inhibitory role in regulating the replication of PRRSV GSWW15 strain in Marc-145 cells.关键词
PRRSV/LncRNATCONS_00179042/Marc-145细胞/过表达/复制Key words
porcine reproductive and respiratory syndrome virus (PRRSV)/LncRNA TCONS_00179042/Marc-145 cell/overexpression/replication分类
农业科技引用本文复制引用
甘利鹏,李平花,李冬,刘在新,卢曾军,张婧,孙普,白伟杰,王治家,曹轶梅,李坤,白兴文,包慧芳..LncRNA TCONS_00179042对Marc-145细胞中PRRSV复制的影响[J].中国兽医科学,2018,48(5):537-544,8.基金项目
中国农业科学院基本科研业务费和创新工程经费资助 （）
