Johann Wolfgang Goethe University, Institute for Molecular Biosciences, Max-von-Laue-Strasse 9, 60438 Frankfurt a.M., Germany.
Mol Microbiol. 2011 May;80(3):666-77. doi: 10.1111/j.1365-2958.2011.07600.x. Epub 2011 Mar 16.
Haloferax volcanii is highly polyploid and contains about 20 copies of the major chromosome. A heterozygous strain was constructed that contained two different types of genomes: the leuB locus contained either the wild-type leuB gene or a leuB:trpA gene introduced by gene replacement. As the trpA locus is devoid of the wild-type trpA gene, growth in the absence of both amino acids is only possible when both types of genomes are simultaneously present, exemplifying gene redundancy and the potential to form heterozygous cells as one possible evolutionary advantage of polyploidy. The heterozygous strain was grown (i) in the presence of tryptophan, selecting for the presence of leuB, (ii) in the presence of leucine selecting for leuB:trpA and (iii) in the absence of selection. Both types of genomes were quantified with real-time PCR. The first condition led to a complete loss of leuB:trpA-containing genomes, while under the second condition leuB-containing genomes were lost. Also in the absence of selection gene conversion led to a fast equalization of genomes and resulted in homozygous leuB-containing cells. Gene conversion leading to genome equalization can explain the escape from 'Muller's ratchet' as well as the ease of mutant construction using polyploid haloarchaea.
火球菌是高度多倍体,含有大约 20 份主要染色体。构建了一个杂合菌株,该菌株含有两种不同类型的基因组:leuB 基因座含有野生型 leuB 基因或通过基因替换引入的 leuB:trpA 基因。由于 trpA 基因座缺失了野生型 trpA 基因,因此只有当两种类型的基因组同时存在时,才能在没有两种氨基酸的情况下生长,这证明了基因冗余和形成杂合细胞的潜力是多倍体的一个可能的进化优势。该杂合菌株在(i)存在色氨酸的情况下生长,选择存在 leuB,(ii)在存在亮氨酸的情况下选择 leuB:trpA,(iii)在没有选择的情况下生长。使用实时 PCR 对两种类型的基因组进行了定量。第一种情况导致含有 leuB:trpA 的基因组完全丢失,而在第二种情况下含有 leuB 的基因组丢失。即使在没有选择的情况下,基因转换也导致基因组快速均等化,并导致纯合 leuB 含有细胞。导致基因组均等化的基因转换可以解释从“Muller's ratchet”逃脱以及使用多倍体盐杆菌轻松构建突变体的原因。