Katinka M D, Bourgain F M
Laboratoire de Génétique Moléculaire, Ecole Normale Supérieure, Paris, France.
EMBO J. 1992 Feb;11(2):725-32. doi: 10.1002/j.1460-2075.1992.tb05105.x.
DNA molecules injected into the macronucleus of Paramecium primaurelia replicate either as free linear telomerized or chromosome integrated molecules. In the present study we show that when a 1.77 kb BamHI DNA fragment harbouring the his3 gene of Saccharomyces cerevisiae was microinjected into the macronucleus, a fraction of the molecules are integrated into the chromosome via an illegitimate recombination process. The injected molecules were mostly inserted at their extremities at multiple points in the genome by replacing the Paramecium sequences. However, insertion sites were not totally at random. Roughly 30% of the molecules were integrated next to or in telomeric repeats. These telomeric repeats were not at the extremities of chromosomes but occupy an internal or interstitial position. We argue that such sites are hotspots for integration as the probability of random insertion near or in an interstitial telomeric site, of which there are 25-60 in a macronucleus is between 5 x 10(-4) and 3 x 10(-5).
注入到双小核草履虫大核中的DNA分子,要么作为游离的线性端粒化分子进行复制,要么作为染色体整合分子进行复制。在本研究中,我们发现,当将携带酿酒酵母his3基因的1.77 kb BamHI DNA片段显微注射到双小核草履虫的大核中时,一部分分子会通过非同源重组过程整合到染色体中。注入的分子大多通过取代双小核草履虫的序列,在基因组的多个位点的末端插入。然而,插入位点并非完全随机。大约30%的分子整合在端粒重复序列旁边或之中。这些端粒重复序列不在染色体末端,而是占据内部或间隙位置。我们认为,这样的位点是整合热点,因为在大核中有25 - 60个间隙端粒位点,在间隙端粒位点附近或之中随机插入的概率在5×10⁻⁴到3×10⁻⁵之间。