A dispensable Plasmodium locus for stable transgene expression.

A dispensable Plasmodium locus for stable transgene expression.
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DOI:
10.1016/j.molbiopara.2009.12.009
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发表时间:
2010-05
影响因子:
1.5
通讯作者:
Kappe, Stefan H. I.
Kappe, Stefan H. I.
中科院分区:
医学4区
文献类型:
--
作者:
Jacobs-Lorena, Vanessa Y.;Mikolajczak, Sebastian A.;Labaied, Mehdi;Vaughan, Ashley M.;Kappe, Stefan H. I.

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核糖体小亚基在伯氏疟原虫和约氏疟原虫中的转基因表达已被用于转基因表达,但这一策略采用的是单交叉整合,因此容易被质粒切除逆转。以核糖体亚单位为靶点也可能对蚊子卵囊的发育产生负面影响。在伯氏假单胞菌中,P230 Paralog基因座已被用于转基因表达。在这里,我们证明了约氏疟原虫S1基因座(子孢子表达基因1)(PY05712)是必不可少的,可以用于在寄生虫整个生命周期中稳定的转基因表达。与约氏疟原虫野生型相比,约氏疟原虫S1−寄生虫在血液期复制、卵囊形成、子孢子产生或肝期发育方面没有缺陷。此外,我们还证明了荧光转基因可以从该位点稳定表达。这表明,S1基因座可以用于在啮齿动物疟疾寄生虫中稳定表达外源基因。
The ribosomal small subunit locus has been used for transgene expression in the rodent malaria parasites, Plasmodium berghei and P. yoelii, but this strategy utilizes single crossover integration and is thus prone to reversion by plasmid excision. Targeting of the ribosomal subunit locus may also have a negative effect on oocyst development in the mosquito. In P. berghei, the p230 paralog locus has been used for transgene expression. Here, we show that the P. yoelii S1 locus (Sporozoite expressed gene 1) (PY05712) is dispensable and can be used for stable transgene expression throughout the parasite life cycle. P. yoelii s1− parasites show no defect in blood stage replication, oocyst formation, sporozoite production, or liver stage development when compared to P. yoelii wildtype parasites. Further, we show that a fluorescent transgene can be stably expressed from this site. This demonstrates that the S1 locus can be utilized for stable expression of heterologous genes in rodent malaria parasites.
DOI: 10.1146/annurev.micro.091208.073403
发表时间: 2009
影响因子: 10.5
作者:
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