TY - JOUR
T1 - Two low-temperature-inducible Chlorella genes for Δ12 and ω-3 Fatty Acid Desaturase (FAD)
T2 - Isolation of Δ12 and ω-3 fad cDNA clones, expression of Δ12 fad in Saccharomyces cerevisiae, and expression of ω-3 fad in Nicotiana tabacum
AU - Suga, Koushirou
AU - Honjoh, Ken-ichi
AU - Furuya, Naoki
AU - Shimizu, Hideyuki
AU - Nishi, Koutarou
AU - Shinohara, Fuminori
AU - Hirabaru, Yoshie
AU - Maruyama, Isao
AU - Miyamoto, Takahisa
AU - Hatano, Shoji
AU - Iio, Masayoshi
N1 - Funding Information:
This research was supported by a Marino forum 21 subsidy and a Grant-in-Aid for Scientiˆc Research (no. 08660164 and 13660127) from the Ministry of Education and Culture of Japan. The authors are grateful to Professor Norio Murata, National Institute of Basic Biology, Okazaki, Japan for kindly providing tobacco plants, to Dr. Y. Ohashi, National Institute of Agrobiological Resources, Tsukuba, Japan for a pBE2113 vector, and to Transgene S.A. for use of the pTG887 plasmid.
PY - 2002/1/1
Y1 - 2002/1/1
N2 - In an attempt to clarify the involvement of fatty acid desaturases (FADs) in the freezing tolerance of Chlorella vulgaris IAM C-27, developed by hardening, we have isolated cDNA clones for two types of FADs from the Chlorella strain, based on the sequence information of genes for Δ12 and ω-3 FADs, respectively desaturating oleic acid (18:1) to linoleic acid (18:2) and linoleic acid (18:2) to linolenic acid (18:3). The deduced amino acid sequence of the first clone, designated CvFad2, showed about 66% similarity to the microsomal Δ12 FADs from several higher plants and this gene had Δ12 FAD activity when expressed in Saccharomyces cerevisiae. The predicted protein encoded by a second gene, designated CvFad3, showed about 60% similarity to the microsomal and plastidial ω-3 FADs from several higher plants. The features of the amino acid sequences of the C- and N-terminal regions of CvFAD3 and fatty acid analysis of polar lipids in transgenic tobacco plant expressing the CvFad3 gene suggested that this gene encodes the microsomal ω-3 FAD. Southern blot analysis showed that both genes were single-copy genes in the genome of the Chlorella strain. Different transcriptional patterns were observed with the two genes during hardening in Northern blot analysis.
AB - In an attempt to clarify the involvement of fatty acid desaturases (FADs) in the freezing tolerance of Chlorella vulgaris IAM C-27, developed by hardening, we have isolated cDNA clones for two types of FADs from the Chlorella strain, based on the sequence information of genes for Δ12 and ω-3 FADs, respectively desaturating oleic acid (18:1) to linoleic acid (18:2) and linoleic acid (18:2) to linolenic acid (18:3). The deduced amino acid sequence of the first clone, designated CvFad2, showed about 66% similarity to the microsomal Δ12 FADs from several higher plants and this gene had Δ12 FAD activity when expressed in Saccharomyces cerevisiae. The predicted protein encoded by a second gene, designated CvFad3, showed about 60% similarity to the microsomal and plastidial ω-3 FADs from several higher plants. The features of the amino acid sequences of the C- and N-terminal regions of CvFAD3 and fatty acid analysis of polar lipids in transgenic tobacco plant expressing the CvFad3 gene suggested that this gene encodes the microsomal ω-3 FAD. Southern blot analysis showed that both genes were single-copy genes in the genome of the Chlorella strain. Different transcriptional patterns were observed with the two genes during hardening in Northern blot analysis.
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U2 - 10.1271/bbb.66.1314
DO - 10.1271/bbb.66.1314
M3 - Article
C2 - 12162554
AN - SCOPUS:0036615280
SN - 0916-8451
VL - 66
SP - 1314
EP - 1327
JO - Bioscience, Biotechnology and Biochemistry
JF - Bioscience, Biotechnology and Biochemistry
IS - 6
ER -