Plohl M, Lucijanić-Justić V, Ugarković D, Petitpierre E, Juan C
Department of Organic Chemistry and Biochemistry, Ruder Bosković Institute, Zagreb, Croatia.
Genome. 1993 Jun;36(3):467-75. doi: 10.1139/g93-064.
The chromosomes of Tribolium confusum have conspicuous bulks of pericentromeric constitutive heterochromatin. The amount of heterochromatin measured by C-banding in metaphase chromosomes is estimated to be 40-45%. It is composed of an A + T rich DNA according to the distamycin A/diamidinophenylindol staining of chromosomes. Restriction analysis of isolated T. confusum genomic DNA shows that this species has a satellite DNA that constitutes about 40% of the genome. Cloning and sequencing experiments reveal a monomer length of 158 base pairs and a copy number of 5.77 x 10(5) per haploid genome. Its sequence is A + T rich (73%), with direct and inverted repeats, one of them with a possibility of forming stable cruciform structure. The abundance, monomer length, and the mutation rate are similar to those found in other satellite families from different species of Tenebrionidae, but no sequence homology has been found among them. No retarded mobility of satellite DNA, characteristic for molecules with sequence-induced curvature, has been detected by electrophoresis on nondenaturing polyacrylamide gels. In situ digestions with restriction enzymes and in situ hybridization show that this satellite DNA is located in pericentromeric positions of all chromosomes coinciding with C-bands.
杂拟谷盗的染色体具有明显的着丝粒周围组成型异染色质块。通过中期染色体的C带测量,异染色质的量估计为40 - 45%。根据染色体的偏端霉素A/二脒基苯基吲哚染色,它由富含A + T的DNA组成。对分离的杂拟谷盗基因组DNA的限制性分析表明,该物种有一种卫星DNA,约占基因组的40%。克隆和测序实验揭示其单体长度为158个碱基对,单倍体基因组的拷贝数为5.77×10⁵。其序列富含A + T(73%),有正向和反向重复序列,其中一个有可能形成稳定的十字形结构。其丰度、单体长度和突变率与拟步甲科不同物种的其他卫星家族相似,但它们之间未发现序列同源性。在非变性聚丙烯酰胺凝胶上进行电泳,未检测到卫星DNA的迁移受阻现象,而这种现象是具有序列诱导弯曲的分子所特有的。用限制性内切酶进行原位消化和原位杂交表明,这种卫星DNA位于所有染色体的着丝粒周围位置,与C带一致。