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Title 

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

Authors 

Won-Kyung HongSun-Yeon HeoB R OhChul Ho KimJung Hoon SohnJ W YangA KondoJeong-Woo Seo

Publisher 

Springer Verlag (Germany)

Issue Date 

2013

Citation 

Bioprocess and Biosystems Engineering, vol. 36, no. 9, pp. 1191-1197

Keywords 

AurantiochytriumCycloheximide resistanceGene transformationRibosomal protein

Abstract 

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 Δ12-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.

ISSN 

1615-7591

Link 

http://dx.doi.org/10.1007/s00449-012-0846-6

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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