Everett J G, Gallie D R
Department of Biochemistry, University of California, Riverside 92521-0129.
Yeast. 1992 Dec;8(12):1007-14. doi: 10.1002/yea.320081203.
An efficient delivery method for introducing in vitro synthesized RNA into yeast has been developed using electroporation. Spheroplast preparation, electroporation, and subsequent expression analysis can be accomplished within a single day. The use of introduced mRNA constructs avoids any complications due to nuclear regulation and is particularly suited for cytoplasmic regulatory studies. Moreover, this technique is useful for introducing those RNAs that cannot be made in vivo, such as poly(A)- mRNAs or RNAs with base modifications. We demonstrate that the Escherichia coli GUS gene and the firefly Luc gene are both excellent reporter genes for RNA electroporation.
利用电穿孔技术，已开发出一种将体外合成的RNA导入酵母的高效递送方法。原生质球制备、电穿孔及后续表达分析可在一天内完成。引入的mRNA构建体的使用避免了由于核调控引起的任何复杂性，特别适用于细胞质调控研究。此外，该技术对于引入那些无法在体内合成的RNA很有用，例如聚腺苷酸化缺陷型mRNA或具有碱基修饰的RNA。我们证明大肠杆菌GUS基因和萤火虫Luc基因都是用于RNA电穿孔的优良报告基因。
