Tagawa M, Omata M, Okuda K
Virology. 1986 Jul 30;152(2):477-82. doi: 10.1016/0042-6822(86)90151-0.
Sequential analysis of duck hepatitis B virus (DHBV) DNA and RNA transcripts in the liver of the early stage of infection was carried out using 1-day-old ducklings inoculated with DHBV-positive serum. Using Southern blot analysis, a band of supercoiled DHBV DNA was detectable at 6 hr, when DHBV-specific poly[A]+RNA was also observed by Northern blot analysis as a faint smear below 3-kb. Bands of RNA at 3.5, 2.7, and 2.5 kb were detected at 12 hr, just before single-stranded viral DNA was detected. A prominent increase in the amount of viral RNA was demonstrated (between 12 hr and 3 days) prior to the increase of DHBV DNA (between 3 and 6 days). Our results suggest that 3.5-kb DHBV-specific RNA synthesized from supercoiled DNA may act as a template of reverse transcription, and that all steps of the replication pathway proposed by Summers and Mason (1982, Cell, 29, 403-415) are completed during 3 days.
利用接种了鸭乙肝病毒(DHBV)阳性血清的1日龄雏鸭,对感染早期肝脏中的DHBV DNA和RNA转录本进行了序列分析。通过Southern印迹分析,在6小时时可检测到超螺旋DHBV DNA条带,此时通过Northern印迹分析也观察到DHBV特异性多聚腺苷酸加尾RNA(poly[A]+RNA),呈3 kb以下的微弱条带。在12小时时检测到3.5、2.7和2.5 kb的RNA条带,就在检测到单链病毒DNA之前。在DHBV DNA增加(3至6天之间)之前(12小时至3天之间),病毒RNA的量显著增加。我们的结果表明,由超螺旋DNA合成的3.5 kb DHBV特异性RNA可能作为逆转录模板,并且Summers和Mason(1982年,《细胞》,第29卷,403 - 415页)提出的复制途径的所有步骤在3天内完成。