Gumpel N J, Rochaix J D, Purton S
Department of Biology, University College London, UK.
Curr Genet. 1994 Nov-Dec;26(5-6):438-42. doi: 10.1007/BF00309931.
The introduction of exogenous DNA into the nuclear genome of Chlamydomonas reinhardtii occurs predominantly via non-homologous (illegitimate) recombination and results in integration at apparently-random loci. Using truncated and modified versions of the C. reinhardtii ARG7 gene in a series of transformation experiments, we demonstrate that homologous recombination between introduced DNA molecules occurs readily in C. reinhardtii, requires a region of homology of no more than 230 bp, and gives rise to intact copies of ARG7 in the nuclear genome. Evidence is presented for homologous recombination between introduced ARG7 DNA and the resident copy of the gene, and for the de-novo synthesis of the ARG7 sequence during transformation.
将外源DNA导入莱茵衣藻的核基因组主要通过非同源(非法)重组发生,并导致在明显随机的位点整合。在一系列转化实验中,我们使用莱茵衣藻ARG7基因的截短和修饰版本,证明引入的DNA分子之间的同源重组在莱茵衣藻中很容易发生,所需的同源区域不超过230 bp,并在核基因组中产生ARG7的完整拷贝。文中提供了引入的ARG7 DNA与该基因的驻留拷贝之间同源重组的证据,以及转化过程中ARG7序列的从头合成的证据。