Geijskes R J, Braithwaite K S, Smith G R, Dale J L, Harding R M
David North Plant Research Centre, Bureau of Sugar Experiment Stations, Indooroopilly, Queensland, Australia.
Arch Virol. 2004 Apr;149(4):791-8. doi: 10.1007/s00705-003-0260-0. Epub 2004 Jan 5.
Sugarcane bacilliform virus (SCBV) DNA molecules larger than the complete genome length of 7.6 kbp were detected in infected plants and in virions. We have confirmed that these high molecular weight nucleic acids were open circular DNA and viral in origin. Due to their open circular conformation, accurate size determination of the DNA molecules was not possible using conventional electrophoresis. Using field inversion gel electrophoresis (FIGE), however, the DNA appeared to increase in genome size increments, with sizes ranging from 1 to 4 genomes (31 kbp) detected. The DNA was packaged into virions, which may explain the observation of purified virions with lengths corresponding to one, two or three times the modal length of 130 nm. The DNA products were possibly concatamers formed during replication as a result of a terminal overlap on the sense strand, and were shown to be overlapped individual genome-length molecules and not covalently-bonded continuous DNA strands. Southern analysis indicated that SCBV sequences are not integrated into the sugarcane genome and that the high molecular weight DNA observed in the sugarcane accessions analysed represents SCBV concatamers.
在受感染的植物和病毒粒子中检测到了大于7.6 kbp完整基因组长度的甘蔗杆状病毒(SCBV)DNA分子。我们已经证实,这些高分子量核酸是开环DNA且起源于病毒。由于其开环构象,使用常规电泳无法准确确定DNA分子的大小。然而,使用脉冲场凝胶电泳(FIGE)时,DNA似乎以基因组大小递增的方式增加,检测到的大小范围为1至4个基因组(31 kbp)。这些DNA被包装进病毒粒子中,这或许可以解释为何观察到纯化的病毒粒子长度对应于130 nm模式长度的1倍、2倍或3倍。这些DNA产物可能是复制过程中由于正义链上的末端重叠而形成的多联体,并且显示为重叠的单个基因组长度分子,而非共价连接的连续DNA链。Southern分析表明,SCBV序列未整合到甘蔗基因组中,在所分析的甘蔗种质中观察到的高分子量DNA代表SCBV多联体。