Rearranging the centromere of the human Y chromosome with phiC31 integrase.
Rearranging the centromere of the human Y chromosome with phiC31 integrase.
复制标题
DOI：
10.1093/nar/gki922
复制
发表时间：
2005
影响因子：
14.9
通讯作者：
Brown, WRA
中科院分区：
文献类型：
--
作者：
Malla, S;Dafhnis-Calas, F;Brookfield, JFY;Smith, MCM;Brown, WRA
我们研究了链霉菌φC31’噬菌体整合酶在鸡DT40杂交体细胞中删除或逆转人Y染色体着丝粒周围1mb DNA的能力。在54%表达整合酶的细胞中观察到互惠和保守位点特异性重组。这些位点在剩下的细胞中无法重组，因为这些位点已经被破坏了。损伤位点的序列表明，损伤是由宿主细胞途径修复重组中间体引起的。重组中间体的损伤倾向与已知的丝氨酸重组酶反应机制是一致的。染色体重排产物的结构与已公布的Y染色体序列一致，表明位点之间高度重复区域的组装精确到约50 kb的分辨率。缺少着丝粒的小染色体没有恢复，这也表明新着丝粒的形成在脊椎动物体细胞中很少发生。φC31整合酶atb位点与鸡基因组未见异位重组。
We have investigated the ability of the integrase from the Streptomyces φC31 ‘phage to either delete or invert 1 Mb of DNA around the centromere of the human Y chromosome in chicken DT40 hybrid somatic cells. Reciprocal and conservative site-specific recombination was observed in 54% of cells expressing the integrase. The sites failed to recombine in the remaining cells because the sites had been damaged. The sequences of the damaged sites indicated that the damage arose as a result of repair of recombination intermediates by host cell pathways. The liability of recombination intermediates to damage is consistent with what is known about the mechanism of serine recombinase reactions. The structures of the products of the chromosome rearrangements were consistent with the published sequence of the Y chromosome indicating that the assembly of the highly repeated region between the sites is accurate to a resolution of about 50 kb. Mini-chromosomes lacking a centromere were not recovered which also suggested that neo-centromere formation occurs infrequently in vertebrate somatic cells. No ectopic recombination was observed between a φC31 integrase attB site and the chicken genome.
登录
查看更多内容
DOI：
10.1073/pnas.95.10.5505
发表时间：
1998-05-12
影响因子：
11.1
作者：
Thorpe, HM;Smith, MCM
通讯作者：
Smith, MCM
影响因子：
56.9
作者：
Schueler, MG;Higgins, AW;Willard, HF
通讯作者：
Willard, HF
影响因子：
64.8
作者：
Guo, F;Gopaul, DN;VanDuyne, GD
通讯作者：
VanDuyne, GD
影响因子：
5.3
作者：
Thyagarajan, B;Olivares, EC;Calos, MP
通讯作者：
Calos, MP
影响因子：
16
作者：
Ghosh, P;Kim, AI;Hatfull, GF
通讯作者：
Hatfull, GF