Insua A, Freire R, Ríos J, Méndez J
Departamento de Biología Celular y Molecular, Universidade da Coruña, Spain.
Chromosome Res. 2001;9(6):495-505. doi: 10.1023/a:1011636714052.
The 5S ribosomal DNA of the mussels Mytilus galloprovincialis and M. edulis was amplified by PCR using contiguous primers. The most general 5S rDNA amplification pattern consisted of several products in both mussels. Two main PCR products of about 250 bp and 760 bp were cloned and sequenced, revealing two classes of 5S rDNA units. These were characterized as containing an identical coding region of 119 bp but with highly divergent spacers. Clones of each unit type exhibited minimal differences except those of the large unit of M. edulis. The sequences analysed of the two mussels possess the same coding region and only six fixed base changes on the spacers. FISH, carried out with specific probes, consistently showed hybridization signals on the largest metacentric pair (two differentiated sites) and with variable frequency on two other metacentric pairs (one site on each pair). Differences in the 5S rDNA distribution between both mussels were not found. In M. edulis, chromosomes carrying 18S-28S rDNA were also identified by FISH. These correspond to two submetacentric-subtelocentric pairs, as was previously reported in M. galloprovincialis, demonstrating that the two rDNA multigene families are located on different chromosome pairs in these mussels.
利用连续引物通过聚合酶链反应(PCR)扩增了地中海贻贝(Mytilus galloprovincialis)和紫贻贝(M. edulis)的5S核糖体DNA。在这两种贻贝中,最常见的5S rDNA扩增模式都由几种产物组成。克隆并测序了约250 bp和760 bp的两个主要PCR产物,揭示了两类5S rDNA单元。这些单元的特征是含有一个119 bp的相同编码区,但间隔区高度不同。除紫贻贝大单元的克隆外,每种单元类型的克隆之间差异极小。对这两种贻贝分析的序列具有相同的编码区,间隔区仅存在六个固定的碱基变化。用特异性探针进行的荧光原位杂交(FISH)始终显示在最大的中着丝粒对(两个不同位点)上有杂交信号,在另外两个中着丝粒对(每对一个位点)上有不同频率的信号。未发现两种贻贝之间5S rDNA分布的差异。在紫贻贝中,通过FISH也鉴定出了携带18S - 28S rDNA的染色体。这些染色体对应于两个亚中着丝粒 - 亚端着丝粒对,正如之前在地中海贻贝中报道的那样,表明这两个rDNA多基因家族位于这些贻贝的不同染色体对上。