急求帮忙翻译!!!(6)

来源：百度知道 编辑：UC知道 时间：2024/06/02 01:23:25
Viruses, macrophage cell cultures, infection, mRNA labeling, and hybridization.
ASFV Pr4 was isolated from ticks obtained from Kruger National Park,
Republic of South Africa, in 1996 (South Africa, Pretoriuskop/96/4) and has had
limited passages in swine macrophage cell cultures (1). Pr4 deletion mutant
Pr4△35, lacking six MGF360 and two MGF530 genes, was constructed as previously
described (54). Macrophage cell cultures were prepared as previously
described (19). Cultures were infected with the Pr4 and Pr4△35 viruses at an
MOI of 10. A virus-depleted, Pr4△35-infected culture supernatant was prepared
to determine if viral infection is required for induction of the macrophage
transcriptional response (MOI of 0.0004). Virus was removed by filtration
through a 0.4-μm-pore-size filter, followed by a 0.1-μm-pore-size low-proteinbinding
filter (Millipore) (prefiltration titer, 1.5*10的8次方; postfiltration titer, 2.9*10的2次方). At 3 and 6 hpi

病毒,巨噬细胞的感染,基因标记及杂交. asfv pr4孤立于蜱所得的克鲁格国家公园,是南非共和国于1996年(南非 pretoriuskop/96/4 )已在有限的机票,在猪巨噬细胞培养物( 1 ) . pr4缺失pr4△35强,缺乏六个mgf360两mgf530基因,构建如前所述( 54 ) . 巨噬细胞的制备如前所述( 19 ) . 文化也受到感染,与pr4和pr4△35病毒在莫伊10 . 病毒枯竭, pr4△35 -病毒培养上清准备,以确定是否感染病毒需诱导巨噬细胞的转录响应( moi 为0.0004 ) . 病毒被拆除,经过滤通过一个0.4μm-孔径大小过滤,然后按0.1 -μm-孔径低proteinbinding滤波器( millipore ) ( prefiltration滴度1.5 * 10的8次方; postfiltration滴度2.9 *10的第2次方) . 在3日和6裴氏,细胞RNA与trizol ( invitrogen ) ,其次是--LiCl沉淀, 大约10μg总RNA标记cy3和cy5的aminoallyl的cDNA标记试剂盒( ambion ) . 幻灯片prehybridized为30分钟, 2 * ssc ( 1 *食糖是0.15米盐加上0.015米的柠檬酸钠) 0.1 %十二烷基硫酸钠( SDS ) -1 %牛血清白蛋白在55 ° C , llowed由两个5分钟洗澡室温( RT )和2 * SSC和0.2 *食糖. cdnas were hybridized to the cDNA阵列为72小时,在45 ° C的杂交cmt庭(康宁公司)与40% formamide - 4 * ssc - 1% sds - 2 * denhardt解- 80 ng聚(一) ( Pharmacia公司) ,每μl( 26岁, 43 )及洗先后在0.2% sds在55 ° C ( 15分钟) , 2 * ssc常温( 15分钟) , d 0.2 * ssc常温( 15分钟) .