Functional Identification of the Plasmodium Centromere and Generation of a Plasmodium Artificial Chromosome
Functional Identification of the Plasmodium Centromere and Generation of a Plasmodium Artificial Chromosome
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DOI：
10.1016/j.chom.2010.02.010
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发表时间：
2010-03-18
影响因子：
30.3
通讯作者：
Waters, Andrew P.
中科院分区：
文献类型：
--
作者：
Iwanaga, Shiroh;Khan, Shahid M.;Waters, Andrew P.
人工染色体是基因转移的有用工具，无论是作为克隆载体还是在染色体生物学研究中都是如此。为了产生疟原虫人工染色体(PAC)，我们必须首先从功能上鉴定和表征该寄生虫的着丝粒。根据疟原虫基因-共线图谱，从伯氏疟原虫5号染色体克隆了一个可能的着丝粒基因(Pbcen5)。在血液期感染期间，含有pbcen5的质粒稳定地保持在寄生虫中，分离效率高，没有药物压力。在蚊子寄生虫的减数分裂和有丝分裂过程中，含有pbcen5的质粒也保持稳定。将pBCen5和端粒整合到一个质粒中，得到线形PAC(L-PAC)。L-PAC分离效率高，并在寄生虫的整个生活史中稳定地保持为一或两个副本。这些结果表明，L-PAC的行为类似于疟原虫染色体，可以作为一种实验研究工具。
The artificial chromosome represents a useful tool for gene transfer, both as cloning vectors and in chromosome biology research. To generate a Plasmodium artificial chromosome (PAC), we had to first functionally identify and characterize the parasite's centromere. A putative centromere (pbcen5) was cloned from chromosome 5 of the rodent parasite P. berghei based on a Plasmodium gene-synteny map. Plasmids containing pbcen5 were stably maintained in parasites during a blood-stage infection with high segregation efficiency, without drug pressure. pbcen5-containing plasmids were also stably maintained during parasite meiosis and mitosis in the mosquito. A linear PAC (L-PAC) was generated by integrating pbcen5 and telomere into a plasmid. The L-PAC segregated with a high efficiency and was stably maintained throughout the parasite's life cycle, as either one or two copies. These results suggest that L-PAC behaves like a Plasmodium chromosome, which can be exploited as an experimental research tool.