期刊论文

Regulation effect of porcine Cyclin A and CDK2 on proliferation of porcine parvovirus

中文

西北农林科技大学学报(自然科学版)

1671-9387

2020

48

8

19-27

10.13207/j.cnki.jnwafu.2020.08.003

31101837:国家自然科学基金 17B041:湖南省教育厅优秀青年项目 ZXXQ201701:衡阳师范学院校企合作项目 18H03:衡阳师范学院校企合作项目 16D20:***专项

本校为第一机构

【目的】探究猪细胞周期素Cyclin A（pCyclinA）及其依赖性激酶2（pCDK2）对猪细小病毒（PPV）增殖的调控作用,为开展PPV复制机制研究提供依据。【方法】采用RT-PCR扩增pCyclinA和pCDK2基因,克隆至真核表达载体pEGFP-C1中构建重组载体,转染猪睾丸细胞（ST）,经新霉素（G418）筛选获得pCyclinA或pCDK2过表达的ST细胞株。设计pCyclinA和pCDK2基因特异性干扰片段（shRNA）CycA894和CDK1163,构建干扰表达载体,转染ST细胞,经G418筛选得到pCyclinA和pCDK2低表达的ST细胞株,实时荧光定量PCR检测其干扰效率。流式细胞术分析pCyclinA和pCDK2过表达和低表达对ST细胞周期的影响。将猪细小病毒NY毒株（PPV-N...

【Objective】This study investigated the regulatory effect of porcine Cyclinin A(pCyclinA)and its dependent kinase 2(pCDK2)on porcine parvovirus(PPV)proliferation,providing the basis for the study off PPV replication mechanism.【Method】The pCyclinAand pCDK2genes were amplified by reverse transcription-polymerase chain reaction(RT-PCR),and then cloned into the eukaryotic expression vector pEGFP-C1.The constructed recombinant vectors were then transfected into porcine testicular cells(ST).Neomycin(G418)was used to screen overexpress ST cell line...