Characterization of heterotrophic transformation algal strains of Dunaliella salina
(0) Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically derived energy. In this study, we attempted to develop a trophic conversion of Dunaliella salina (D. salina), namely from an obligate photoautotrophic to heterotrophic by a gene encoding glucose transporter (Glut1 ). The inducible heterotrophic expression vector pMDDGN-Bar and constitutive heterotrophic expression vector G5Glut1-Bar were transformed into cells of D. salina through optimized electroporation method, respectively, and then heterotrophic algal strains of D. salina were selected using the PPT liquid and solid screening procedures. All the wild type cells of the alga died when PPT was used at the concentration of 6 μg/mL, while transformed cells of D. salina grew well. The three constitutive heterotrophic algal strains and 2 inducible heterotrophic algal strains were named as C1, C2, C3, I1 and I2, respectively. In addition, the results of RT-PCR showed that 5 strains of transformants all appeared about 250 bp of specific DNA bands, suggesting that foreign Glut1 gene had been integrated successfully into the genome of heterotrophic strains of D. salina. Further, Southern blots were utilized to analyze the transgene copy number of heterotrophic transformation algal strains of D. salina. The results revealed that the copy numbers of C1, C2, C3 I1 and I2 were 2, 1, 3, 3 and 2, respectively. These findings may lay a foundation for developing transgenic D. salina bioreactor.
Dunaliella salina electroporation transformation Southern blots copy number
Wenbing Lv Hongtao Liu Lexun Xue
Laboratory for Cell Biology, Department of Biology, Zhengzhou University, Zhengzhou, 450001, China
国际会议
南京
英文
200-203
2010-05-12（万方平台首次上网日期，不代表论文的发表时间）