莱茵衣藻血红素加氧酶1过表达载体的构建和遗传转化

Expression Vector Construction of Heme Oxygenase-1 from Chlamydomonas reinhardtii and Transformation

莱茵衣藻(Chlamydomonas reinhardti)是一种3套基因组都能进行遗传转化的真核生物,作为一种模式生物,它被用于生物学研究的各个领域。目前,发现血红素加氧酶具有多种功能活性,但关于它的作用机理还不是很清楚。本研究利用分子生物学技术,构建了莱茵衣藻HO-1过表达载体,用SpeI和BglII双酶切,DNA测序,GUS染色,PCR检测证明表达载体构建成功。将此构建通过农杆菌介导法导入莱茵衣藻细胞中,获得了能够稳定遗传的转化子。上述结果为后续进一步的功能研究奠定基础。

The green alga Chlamydomonas reinhardtii,a unicellular eukaryote has three genomes. It is a valuable model system for biological study. Recently,heme oxygenase-1 （HO-1）is shown to play multiple roles in biological processes,but the accurate functions of its gene are largely unknown. In this study, we constructed an expression vector harboring HO-1 from Chlamydomonas reinhardtii. Digestion with Spe I and Bgl II, DNA sequencing,GUS staining and PCR detecting proved that the HO-1 gene has been cloned into the expression vector. The constructs were then transferred into the cells by Agrobacterium-mediated method. RT-PCR results indi- cated that the reorganized vector has worked well in the cells and expressed a great quantity of HO-1 products. The result of the present study lays the foundation for the further research on function.