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DNA探针在盘基网柄菌野生分离株分类学研究中的应用。

The use of DNA probes for taxonomic study of dictyostelium wild isolates.

作者信息

Evans W B, Hughes J E, Welker D L

机构信息

Department of Biology, Utah State University, Logan 84322-5500.

出版信息

Genetics. 1988 Jul;119(3):561-9. doi: 10.1093/genetics/119.3.561.

Abstract

The classification of 27 wild isolates assigned to Dictyostelium discoideum on the basis of morphological criteria was reexamined using probes specific for DNA sequences cloned from the type strain NC4. These probes included ones specific for ribosomal spacer DNA regions and for a ribosomal RNA coding sequence, as well as probes for two chromosomal gene families (actin and discoidin) and for the DIRS-1 transposable element. Four isolates (AC4, WS526, WS584 and ZA3A) which had previously been shown to have unusual mating characteristics were distinctly different from other isolates. We interpret these differences as indicating that the four atypical isolates represent species other than D. discoideum. Probes for the ribosomal spacer DNA either did not hybridize to the DNA of these four isolates or had decreased levels of hybridization to EcoRI restriction fragments of different lengths to that observed with the type strain. With the discoidin probe, all isolates had DNA fragments that hybridized but AC4, WS526, WS584 and ZA3A lacked a pair of fragments that were conserved in NC4 and other isolates. With the actin probe, AC4, WS526, WS584 and ZA3A lacked numerous fragments that the other isolates shared with NC4. The DIRS-1 probes showed strong hybridization with ZA3A and weak hybridization to the other three isolates; however, the major EcoRI fragment in WS526 and WS584 was smaller than that in NC4 while ZA3A and AC4 had fragments of similar size to that in NC4.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

基于形态学标准被归类为盘基网柄菌的27个野生分离株,使用从模式菌株NC4克隆的DNA序列特异性探针进行了重新检测。这些探针包括核糖体间隔区DNA区域和核糖体RNA编码序列的特异性探针,以及两个染色体基因家族(肌动蛋白和盘状菌素)和DIRS-1转座元件的探针。先前已证明具有异常交配特征的四个分离株(AC4、WS526、WS584和ZA3A)与其他分离株明显不同。我们将这些差异解释为表明这四个非典型分离株代表了不同于盘基网柄菌的物种。核糖体间隔区DNA探针要么不与这四个分离株的DNA杂交,要么与不同长度的EcoRI限制性片段的杂交水平低于模式菌株。使用盘状菌素探针时,所有分离株都有杂交的DNA片段,但AC4、WS526、WS584和ZA3A缺少在NC4和其他分离株中保守的一对片段。使用肌动蛋白探针时,AC4、WS526、WS584和ZA3A缺少其他分离株与NC4共有的许多片段。DIRS-1探针与ZA3A显示出强杂交,与其他三个分离株显示出弱杂交;然而,WS526和WS584中的主要EcoRI片段比NC4中的小,而ZA3A和AC4具有与NC4中大小相似的片段。(摘要截短于250字)

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