Zhang P, Spradling A C
Howard Hughes Medical Institute Research Laboratories, Carnegie Institution of Washington, Baltimore, Maryland 21210.
Genetics. 1993 Feb;133(2):361-73. doi: 10.1093/genetics/133.2.361.
We have investigated how Drosophila P element insertions are distributed in the chromosomal region near their starting site. A single P element residing in the euchromatin of minichromosome Dp1187 was mobilized following a cross to the delta 2-3 (99B) strain, and progeny bearing transpositions were identified with a minimum of bias by performing Southern blots on progeny. Approximately 1-2% of all progeny minichromosomes contained new insertions. Many of these "local transpositions" landed very close to or within the starting P element; however, nearly 1% of all progeny chromosomes contained new insertions 1-180 kb from the donor element. More local insertions were observed in the progeny of females than from male parents, and most occurred in a preferred orientation relative to the starting element. These observations suggested that donor elements are frequently excised and reinserted locally without ever dissociating from a transposition complex. The high frequency and diverse distribution of local transpositions recovered from females suggested that the efficiency of insertional mutagenesis can be significantly enhanced by using a starting P element(s) located near the target of interest.
我们研究了果蝇P因子插入在其起始位点附近的染色体区域是如何分布的。位于小染色体Dp1187常染色质中的单个P因子在与delta 2-3(99B)品系杂交后被激活,通过对后代进行Southern杂交,以最小的偏差鉴定出携带转座的后代。所有后代小染色体中约有1%-2%含有新的插入。许多这些“局部转座”落在起始P因子非常靠近或在其内部的位置;然而,所有后代染色体中近1%含有距供体因子1-180 kb的新插入。在雌性后代中观察到的局部插入比雄性亲本的更多,并且大多数相对于起始因子以优先方向发生。这些观察结果表明,供体因子经常被切除并在局部重新插入,而从未从转座复合体中解离。从雌性中回收的局部转座的高频率和多样分布表明,通过使用位于感兴趣靶标附近的起始P因子,可以显著提高插入诱变的效率。