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芝田硫化叶菌（Sulfolobus shibatae）是一种极端嗜热古菌,其所携带的温和病毒SSV1是研究古菌DNA复制的一个非常合适的系统.杂交分析表明,静置培养时,芝田硫化叶菌细胞内几乎检测不到游离SSV1 DNA,病毒 DNA主要以整合状态存在.经紫外或丝裂霉素诱导后,细胞内游离SSV1 DNA含量明显增加,但整合SSV1 DNA拷贝数不变.丝裂霉素诱导作用的重复性明显高于紫外诱导.另外,当细胞由静置培养改为摇床培养时,SSV1 DNA也被诱导复制.据此,建立了 SSV1 DNA复制的丝裂霉素诱导方法.采用此法,芝田硫化叶菌细胞内游离 SSV1 DNA含量在诱导后 10h开始明显增加,并在12～15h达到最大值.诱导后细胞内游离病毒 DNA的拷贝数可达到 50.上述诱导方法的建立以及对诱导后SSV1 DNA复制行为的描述为确定 SSV1病毒 DNA的复制原点、分析其复制模式奠定了基础.
Sulfolobus shibatae is an extremely thermophilic archaea, and its mild virus SSV1 is a very suitable system for studying the DNA replication of archaea. Hybridization analysis showed that during the stationary culture, Almost no intracellular free SSV1 DNA was detected, and the viral DNA mainly existed in the integrated state.Under ultraviolet or mitomycin-induced cell-free SSV1 DNA content increased significantly, but the integrated SSV1 DNA copy number unchanged.Methymidine In addition, SSV1 DNA was also induced to replicate when the cells were changed from resting to shaker cultures.Therefore, an induction method of mitomycin SSV1 DNA replication was established by using In this method, the content of free SSV1 DNA in S. cuneata cells increased significantly at 10h after induction and reached the maximum at 12 ~ 15h, and the number of free viral DNA after induction was 50. The establishment of the induction method and Describing the SSV1 DNA replication behavior after induction establishes the basis for determining the replication origin of the SSV1 viral DNA and analyzing its replication pattern.