Nuclear gene targeting in Chlamydomonas reinhardtii

通过模型基因，我们已经表明，对于莱茵衣藻，单链DNA能够比双链DNA更有效地靶向或修饰核基因。此外，非同源DNA整合几乎被完全抑制。这些发现开启了在绿色藻类中沉默或修饰任何选定基因的可能性。在接下来的三年里，我们打算优化这种“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，我们将过量表达来自龟裂链霉菌Ps.铜绿假单胞菌或衣原体本身或在转化期间将纯化的重组蛋白添加到ssDNA。

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.