A transgene expression system for the marine microalgae Aurantiochytrium sp KRS101 using a mutant allele of the gene encoding ribosomal protein L44 as a selectable transformation marker for cycloheximide resistance

作者：Hong Won Kyung; Heo Sun Yeon; Oh Baek Rock; Kim Chul Ho; Sohn Jung Hoon; Yang Ji Won; Kondo Akihiko; Seo Jeong Woo^*

来源：Bioprocess and Biosystems Engineering, 2013, 36(9): 1191-1197.

DOI：10.1007/s00449-012-0846-6

• Aurantiochytrium
• Ribosomal protein
• Cycloheximide resistance
• Gene transformation

In the present study, we established a genetic system for manipulating the oleaginous heterotrophic microalgae Aurantiochytrium sp. KRS101, using cycloheximide resistance as the selectable marker. The gene encoding ribosomal protein L44 (RPL44) of Aurantiochytrium sp. KRS101 was first identified and characterized. Proline 56 was replaced with glutamine, affording cycloheximide resistance to strains encoding the mutant protein. This resistance served as a novel selection marker. The gene encoding the Delta 12-fatty acid desaturase of Mortierella alpina, used as a reporter, was successfully introduced into chromosomal DNA of Aurantiochytrium sp. KRS101 via 18S rDNA-targeted homologous recombination. Enzymatic conversion of oleic acid (C18:1) to linoleic acid (C18:2) was detected in transformants but not in the wild-type strain.

• 出版日期2013-9