粗神经孢子虫转化后对肌醇利用的改进选择

R. L. Metzenberg
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Metzenberg Department of Biological Sciences, Stanford U niversity, Stanford , California 94305-5020; present address, Dept. of Chemistry and Biochemistry, University of California, Los Angeles CA 90095-1569 Plasmids based on the cloned inositol gene of Neurospora crassa are potentially very useful as a transformation marker, or for techniques like insertional mutagenesis. Transformation is generally done on spheroplasts stabilized with sorbitol, or by electroporation of conidia, also suspended in sorb itol. For transformation to inositol independence, however, this gives an unacceptable background of nontransformants that grow on the nominally inositol-free medium. It appears that sorbitol always contains a trace of inositol that cannot be completely removed by recrystallization. It seems possible that sorbitol, which differs from inositol only by a pair of hydrogens, spontaneously generates the latter by air oxidation. 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摘要

基于克隆的粗神经孢子虫肌醇基因的质粒作为转化标记或插入诱变等技术可能非常有用。本作品采用知识共享署名-相同方式共享4.0许可协议。这篇简短的文章可以在真菌遗传学报告中找到:http://newprairiepress.org/fgr/vol49/iss1/9 18真菌遗传学通讯:粗神经孢子虫转化后肌醇利用的改进选择Robert L. Metzenberg,斯坦福大学生物科学系,斯坦福,加州94305-5020;基于克隆的粗神经孢子虫肌醇基因的质粒作为转化标记或插入诱变等技术非常有用。转化通常是在用山梨糖醇稳定的球质体上进行的,或者通过电穿孔的分生孢子,也悬浮在山梨糖醇中。然而,对于转化为肌醇独立性,这给出了在名义上不含肌醇的培养基上生长的非转化体的不可接受的背景。山梨糖醇似乎总是含有不能通过重结晶完全去除的肌醇。山梨糖醇与肌醇只有一对氢的区别,山梨糖醇似乎有可能通过空气氧化自发地产生肌醇。在等摩尔的基础上用a-甲基葡萄糖苷代替山梨糖醇可以避免这个问题。a-甲基葡萄糖苷作为一种渗透剂很好地发挥作用,并为in1突变体向肌醇独立性的转化提供了干净的背景。新草原出版社2017年出版
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Improved selection for inositol-utilization following transformation of Neurospora crassa
Plasmids based on the cloned inositol gene of Neurospora crassa are potentially very useful as a transformation marker, or for techniques like insertional mutagenesis. Creative Commons License This work is licensed under a Creative Commons Attribution-Share Alike 4.0 License. This brief note is available in Fungal Genetics Reports: http://newprairiepress.org/fgr/vol49/iss1/9 18 Fungal Genetics Newsletter Improved selection for inositol-utilization following transformation of Neurospora crassa Robert L. Metzenberg Department of Biological Sciences, Stanford U niversity, Stanford , California 94305-5020; present address, Dept. of Chemistry and Biochemistry, University of California, Los Angeles CA 90095-1569 Plasmids based on the cloned inositol gene of Neurospora crassa are potentially very useful as a transformation marker, or for techniques like insertional mutagenesis. Transformation is generally done on spheroplasts stabilized with sorbitol, or by electroporation of conidia, also suspended in sorb itol. For transformation to inositol independence, however, this gives an unacceptable background of nontransformants that grow on the nominally inositol-free medium. It appears that sorbitol always contains a trace of inositol that cannot be completely removed by recrystallization. It seems possible that sorbitol, which differs from inositol only by a pair of hydrogens, spontaneously generates the latter by air oxidation. Substituting a-methyl glucoside for the sorbitol on an equimolar basis can circumvent this problem. a-Methyl glucoside works well as an osmoticum, and gives clean backgrounds in transformation of inl mutants to inositol independence. Published by New Prairie Press, 2017
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