Utility of the pat gene as a selectable marker gene in production of transgenic Dunaliella salina

Fisheries and Aquatic Sciences

Hyo Sun Jung¹  Dong Soo Kim² 

[1] Center for Risk Assessment of Oceans and Fisheries Living Modified Organisms, Pukyong National University;Department of Marine Bio-Materials and Aquaculture, Pukyong National University;

关键词: Dunaliella salina;    Microalga;    pat gene;    Selectable marker gene;   

DOI  :  10.1186/s41240-016-0030-z

来源: DOAJ

【 摘 要 】

Abstract Background The objective of this study was to develop an efficient selectable marker for transgenic Dunaliella salina. Results Tests of the sensitivity of D. salina to the antibiotic chloramphenicol and the herbicide Basta® showed that cells (1.0 × 106 cells/ml) treated with 1000 or 1500 μg/ml chloramphenicol died in 8 or 6 days, respectively, whereas D. salina cells (1.0 × 106 cells/ml) treated with 5, 10, 20, or 40 μg/ml Basta® died in 2 days. Therefore, D. salina is more sensitive to Basta® than to chloramphenicol. To examine the possibility of using the phosphinothricin N-acetyltransferase (pat) gene as a selectable marker gene, we introduced the pat genes into D. salina with particle bombardment system under the condition of helium pressure of 900 psi from a distance of 3 cm. PCR analysis confirmed that the gene was stably inserted into the cells and that the cells survived in 5 μg/ml Basta®, the medium used to select the transformed cells. Conclusions The findings of this study suggest that the pat gene can be used as an efficient selectable marker when producing transgenic D. salina.

【 授权许可】

Unknown