一种新型的、快速且高效的 Neurospora 转化程序。

A new, rapid and efficient transformation procedure for Neurospora.

机构信息

Departments of Biochemistry and Genetics, The Ohio State University, 43210, Columbus, Ohio, USA.

出版信息

Curr Genet. 1984 Jan;8(1):77-9. doi: 10.1007/BF00405435.

Abstract

A new, rapid, efficient and reliable method for transforming Neurospora crassa is described. In this procedure, germinated conidia are treated with lithium acetate, then incubated with DNA, followed by exposure to polyethylene glycol and then a brief heat shock, prior to plating on selective medium. Optimal conditions to achieve a high transformation rate are reported. Transformation can be obtained with both circular and linear plasmid DNA and also with genomic DNA. Although the rate is substantially decreased, transformation was also obtained with relatively impure DNA preparations, such as that made via rapid "miniprep" procedures. This transformation technique is simple and reliable and provides a considerable savings in time and materials.

摘要

描述了一种新的、快速、高效和可靠的转化粗糙脉孢菌（Neurospora crassa）的方法。在这个过程中，用醋酸锂处理发芽的分生孢子，然后与 DNA 孵育，接着用聚乙二醇处理并进行短暂的热休克，然后在选择性培养基上进行平板培养。报告了达到高转化率的最佳条件。可以用圆形和线性质粒 DNA 以及基因组 DNA 进行转化。虽然转化率大大降低，但也可以用相对不纯的 DNA 制剂进行转化，例如通过快速“小量制备”程序制备的 DNA。这种转化技术简单可靠，在时间和材料上都有很大的节省。