Nuclear gene targeting in Chlamydomonas reinhardtii

通过模型基因，我们已经证明了莱茵衣藻单链DNA比双链DNA更有效地靶向或修饰核基因。此外，非同源dna整合几乎完全被抑制。这些发现开启了沉默或修饰绿藻中任何选定基因的可能性。在接下来的三年中，我们打算优化这个“核基因靶向系统”，使其特别适用于弱表达基因。为了提高重组效率，我们将从链霉菌、铜绿假单胞菌或衣藻本身过表达一些参与重组过程的基因，或者在转化过程中向ssDNA中加入纯化的重组蛋白。

By means of a model gene we have shown for Chlamydomonas reinhardtii that single stranded DNA is able to target or modifiy nuclear genes more efficiently than double stranded DNA. In addition, non-homologous DNA-Integration is nearly completely suppressed. These findings open the possibility of silencing or modification of any selected gene in green algae. During the next three years we intend to optimize this "Nuclear Gene Targeting System" and make it applicable especially for weakly expressed genes. To increase the recombination efficiency we will over-express some genes that are involved in recombination processes from Streptomyces rimosus, Ps. aeruginosa or Chlamydomonas itself or add purified recombination proteins to ssDNA during transformation.