Feng Shuying, Xue Lexun, Liu Hongtao, Lu Pengju
Laboratory for Cell Biology, The First Affiliated Hospital, Zhengzhou University Medical College, Zhengzhou University, 40 Daxue Road, Zhengzhou, Henan, 450052, People's Republic of China.
Mol Biol Rep. 2009 Jul;36(6):1433-9. doi: 10.1007/s11033-008-9333-1. Epub 2008 Sep 16.
Dunaliella salina has been exploited as a new type of bioreactor due to its unique advantages. However, this bioreactor application was restricted for absence of a high-efficiency and stable transformation method at present. In the present study, the cells of D. salina were transformed by glass beads. The results of histochemical staining revealed that the GUS gene was successfully expressed in the positive transformants, and PCR and PCR-Southern blot analysis further demonstrated that the bar gene was integrated into the D. salina genome. Moreover, the three transformation methods, including glass beads, bombardment particle and electroporation, were compared for screening a high-efficiency transformation method for gene engineering of D. salina. The results showed that transformation efficiency of the glass beads was the highest, approximately 10(2) transformants/microg DNA. It is concluded that the established glass beads method has been demonstrated to be an optimal transformation way for D. salina.
盐生杜氏藻因其独特优势已被开发为一种新型生物反应器。然而，由于目前缺乏高效稳定的转化方法，这种生物反应器的应用受到限制。在本研究中，盐生杜氏藻细胞通过玻璃珠进行转化。组织化学染色结果显示，GUS基因在阳性转化体中成功表达，PCR及PCR-Southern杂交分析进一步证明bar基因已整合到盐生杜氏藻基因组中。此外，对玻璃珠法、粒子轰击法和电穿孔法这三种转化方法进行了比较，以筛选出用于盐生杜氏藻基因工程的高效转化方法。结果表明，玻璃珠法的转化效率最高，约为10(2)个转化体/μg DNA。结论是，所建立的玻璃珠法已被证明是盐生杜氏藻的一种最佳转化方法。
