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在其组成核中具有互补重复和缺失的粗糙脉孢菌异核体提供了一种鉴定核有限基因的方法。

Neurospora Heterokaryons with Complementary Duplications and Deficiencies in Their Constituent Nuclei Provide an Approach to Identify Nucleus-Limited Genes.

作者信息

Giri Dev Ashish, Rekha S, Kasbekar Durgadas P

机构信息

Centre for DNA Fingerprinting and Diagnostics, Hyderabad 500001, India Graduate Studies, Manipal University, Manipal 576104, Karnataka, India.

Centre for DNA Fingerprinting and Diagnostics, Hyderabad 500001, India.

出版信息

G3 (Bethesda). 2015 Apr 20;5(6):1263-72. doi: 10.1534/g3.115.017616.

Abstract

Introgression is the transfer of genes or genomic regions from one species into another via hybridization and back-crosses. We have introgressed four translocations (EB4, IBj5, UK14-1, and B362i) from Neurospora crassa into N. tetrasperma. This enabled us to construct two general types of heterokaryons with mat-A and mat-a nuclei of different genotypes: one type is [T + N] (with one translocation nucleus and one normal sequence nucleus), and the other is [Dp + Df] (with one nucleus carrying a duplication of the translocation region and the other being deleted for the translocation region). Self-crossing these heterokaryons again produced [T + N] and [Dp + Df] progeny. From conidia (vegetative spores) produced by the heterokaryotic mycelia, we obtained self-fertile (heterokaryotic) and self-sterile (homokaryotic) derivative strains. [T + N] heterokaryons produced homokaryotic conidial derivatives of both mating types, but [Dp + Df] heterokaryons produced viable conidial homokaryons of only the mating type of the Dp nucleus. All four [T + N] heterokaryons and three [Dp + Df] heterokaryons produced both self-sterile and self-fertile conidial derivatives, but the [Dp(B362i) + Df(B362i)] heterokaryons produced only self-sterile ones. Conceivably, the Df(B362i) nuclei may be deleted for a nucleus-limited gene required for efficient mitosis or nuclear division, and whose deficit is not complemented by the neighboring Dp(B362i) nuclei. A cross involving Dp(EB4) showed repeat-induced point mutation (RIP). Because RIP can occur in self-crosses of [Dp + Df] but not [T + N] heterokaryons, RIP alteration of a translocated segment would depend on the relative numbers of [Dp + Df] vs. [T + N] ancestors.

摘要

基因渗入是指基因或基因组区域通过杂交和回交从一个物种转移到另一个物种。我们已将粗糙脉孢菌的四个易位(EB4、IBj5、UK14 - 1和B362i)导入四孢脉孢菌。这使我们能够构建两种不同基因型的具有mat - A和mat - a核的异核体:一种类型是[T + N](一个易位核和一个正常序列核),另一种是[Dp + Df](一个核携带易位区域的重复,另一个核缺失易位区域)。这些异核体再次自交产生了[T + N]和[Dp + Df]后代。从异核体菌丝体产生的分生孢子(营养孢子)中,我们获得了自育(异核体)和自不育(同核体)的衍生菌株。[T + N]异核体产生了两种交配型的同核体分生孢子衍生物,但[Dp + Df]异核体仅产生了与Dp核交配型相同的可存活同核体分生孢子。所有四个[T + N]异核体和三个[Dp + Df]异核体都产生了自不育和自育的分生孢子衍生物,但[Dp(B362i) + Df(B362i)]异核体仅产生了自不育的衍生物。可以想象,Df(B362i)核可能缺失了高效有丝分裂或核分裂所需的核限基因,且其缺陷不能被相邻的Dp(B362i)核互补。涉及Dp(EB4)的杂交显示出重复诱导点突变(RIP)。因为RIP可发生在[Dp + Df]而非[T + N]异核体的自交中,所以易位片段的RIP改变将取决于[Dp + Df]与[T + N]祖先的相对数量。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b93f/4478554/cbab2b854cf3/1263f1.jpg

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