Pannekoek Yvonne, van der Ende Arie
Department of Medical Microbiology, Academic Medical Center, Center for Infection and Immunity, Amsterdam, The Netherlands.
Methods Mol Biol. 2012;799:73-89. doi: 10.1007/978-1-61779-346-2_5.
A riboregulated network, in which small RNAs (sRNAs) regulate the stability and thus translation of transcripts (mRNA), has only recently been discovered in prokaryotes. Yet, during the last 5 years, hundreds of sRNAs have been identified in various bacterial species by using a wide variety of both computational and experimental approaches. The majority of the sRNAs interact with the 5'-untranslated region (UTR) of target mRNAs, thereby influencing the stability of target mRNAs, or by either suppressing or upregulating the ribosome entry to the mRNAs influencing translation. Here, we describe experimental approaches successfully used in our laboratory to identify and functionally characterize sRNAs in vivo in our model micro-organism Neisseria meningitidis.
一种核糖调节网络,其中小RNA(sRNA)调节转录本(mRNA)的稳定性进而调节其翻译,直到最近才在原核生物中被发现。然而,在过去5年里,通过使用各种各样的计算和实验方法,在各种细菌物种中已经鉴定出了数百种sRNA。大多数sRNA与靶标mRNA的5'非翻译区(UTR)相互作用,从而影响靶标mRNA的稳定性,或者通过抑制或上调核糖体与mRNA的结合来影响翻译。在这里,我们描述了在我们实验室中成功使用的实验方法,用于在我们的模式微生物脑膜炎奈瑟菌体内鉴定sRNA并对其进行功能表征。