Title Establishment of a Micro-Particle Bombardment Transformation System for Dunaliella salina
Author Congping Tan1,2, Song Qin1,*, Qun Zhang1,2, Peng Jiang1, and Fangqing Zhao1,2
Address 1Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, P.R.China, 2Graduate School of Chinese Academy of Sciences, Beijing 100039, P.R.China
Bibliography Journal of Microbiology, 43(4),361-365, 2005,
Key Words Dunaliella salina, micro-particle bombardment, GUS, bar
Abstract In this study, we chronicle the establishment of a novel transformation system for the unicellular marine green alga, Dunaliella salina. We introduced the CaMV35S promoter-GUS construct into D. salina with a PDS1000/He micro-particle bombardment system. Forty eight h after transformation, via histochemical staining, we observed the transient expression of GUS in D. salina cells which had been bombarded under rupture-disc pressures of 450 psi and 900 psi. We observed no GUS activity in either the negative or the blank controls. Our findings indicated that the micro-particle bombardment method constituted a feasible approach to the genetic transformation of D. salina. We also conducted tests of the cells'' sensitivity to seven antibiotics and one herbicide, and our results suggested that 20 ug/ml of Basta could inhibit cell growth completely. The bar gene, which encodes for phosphinothricin acetyltransferase and confers herbicide tolerance, was introduced into the cells via the above established method. The results of PCR and PCR-Southern blot analyses indicated that the gene was successfully integrated into the genome of the transformants.
Download PDF p.361-365.pdf