Heslip T R, Hodgetts R B
Department of Genetics, University of Alberta, Edmonton, Canada.
Genetics. 1994 Dec;138(4):1127-35. doi: 10.1093/genetics/138.4.1127.
Targeted transposition is the replacement of one P element with another. We are exploiting this unique property of P elements to study the complex regulatory domain of the Dopa decarboxylase (Ddc) gene in Drosophila melanogaster. P element constructs targeted to the same site in the genome will be subjected to the same position effect. This allows the subtle effects typical of most mutations in the Ddc regulatory region to be measured in the absence of the variable influences of position effects which are associated with the current method of germline transformation. We have investigated some of the parameters affecting targeted transposition of a Ddc transposon, P[Ddc], into a P element allele at the vestigial locus. These events were detected by an increased mutant vg phenotype. The location of the donor transposon in cis or in trans to the target had little effect on the frequency of targeting. Likewise, the mobility of different donor elements, as measured by their rate of transposition to a different chromosome, varied nearly 20-fold, while the rate of targeted transposition was very similar between them. All targeted alleles were precise replacements of the target P element by P[Ddc], but in several cases the donor was inserted in the opposite orientation. The targeted alleles could be described as the result of a replicative, conversion-like event.
靶向转座是用一个P因子替换另一个P因子。我们正在利用P因子的这一独特特性来研究黑腹果蝇中多巴脱羧酶(Ddc)基因的复杂调控区域。靶向基因组中同一位点的P因子构建体将受到相同的位置效应影响。这使得在没有与当前种系转化方法相关的位置效应的可变影响的情况下,能够测量Ddc调控区域中大多数突变典型的细微效应。我们研究了一些影响Ddc转座子P[Ddc]靶向转座到残翅位点的P因子等位基因的参数。这些事件通过突变的vg表型增加来检测。供体转座子与靶标顺式或反式的位置对靶向频率影响很小。同样,不同供体元件的移动性,以它们转座到不同染色体的速率来衡量,相差近20倍,而它们之间的靶向转座速率非常相似。所有靶向等位基因都是P[Ddc]对靶标P因子的精确替换,但在几种情况下,供体以相反的方向插入。靶向等位基因可以被描述为复制性、类似转换事件的结果。