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Genome-wide screening and transcriptional profile analysis of desaturase genes in the European corn borer moth

Insect Science 2012年第1期19卷 55-63页

作者： Bingye Xue Alejandro P. Rooney Wendell L. Roelofs Department of Entomology NYSAES-Comell University Geneva NY Crop Bioprotection Research Unit National Centerfor Agricultural Utilization Research Agricultural Research Service U.S. Department of Agricuiture Peoria IL USA

Acyl-coenzyme A （Acyl-CoA） desaturases play a key role in the biosynthesis of female moth sex pheromones. desaturase genes are encoded by a large multigene family, and they have been divided into five subgroups on the basis of biochemical functionality and phylogenetic affinity. In this study both copy numbers and transcriptional levels of desaturase genes in the European corn borer （ECB）, Ostrinia nubilalis, were investigated. The results from genome-wide screening of ECB bacterial artificial chromosome （BAC） library indicated there are many copies of some desaturase genes in the genome. An open reading frame （ORF） has been isolated for the novel desaturase gene ECB ezi-A11~ from ECB gland complementary DNA and its functionality has been analyzed by two yeast expression systems. No functional activities have been detected for it. The expression levels of the four desaturase genes both in the pheromone gland and fat body of ECB and Asian corn borer （ACB）, O. furnacalis, were determined by real-time polymerase chain reaction. In the ECB gland, All is the most abundant, although the amount of A14 is also considerable. In the ACB gland, A14 is the most abundant and is 100 times more abundant than all the other three combined. The results from the analysis of evolution of desaturase gene transcription in the ECB, ACB and other moths indicate that the pattern of A 11 gene transcription is significantly different from the transcriptional patterns of other desaturase genes and this difference is tied to the underlying nucleotide composition bias of the genome.

关键词： bacterial artificial chromosome (BAC) library desaturase European corn borer G+C sex pheromone

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Identification and characterization of a delta-12 fatty acid desaturase gene from marine microalgae Isochrysis galbana

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Acta Oceanologica Sinica 2019年第2期38卷 107-113页

作者： Xiaotian Han Shuai Wang Li Zheng Wanshun Liu College of Marine Life Sciences Ocean University of China Changjiang Estuary Marine Ecosystem Research Station Institute of OceanologyChinese Academy of Sciences Marine Ecology and Environmental Science Laboratory Pilot National Laboratory for Marine Science and Technology (Qingdao) Key Laboratory for Marine Bioactive Substances and Modern Analytical Technology First Institute of OceanographyMinistry of Natural Resources

The cDNA of the delta-12 fatty acid desaturase gene, IgFAD2, was cloned from the marine microalgae Isochrysis galbana, a species capable of producing docosahexaenoic acid. Sequence analysis indicated that the open reading frame measured a length of 1 158 bp and encoded 386 amino acids with a predicted molecular weight of 42.8 kDa and an isoelectric point of 9.2. Computational analysis of the protein sequence of IgFAD2 showed typical features of membrane-bound desaturase such as three conserved histidine boxes along with four membranespanning regions that were universally present among plant desaturases. Quantitative real-time PCR results showed that the abundance of IgFAD2 transcript was significantly upregulated under different environmental stresses including low temperature(15℃), high salinity(salinity of 62 and 93), and nitrogen starvation(220 μmol/L). Heterologous expression indicated that yeast cells transformed with a plasmid construct containing IgFAD2 could convert endogenous oleic acid(18:1^(?9), OA) into linoleic acid(18:2^(?9, 12), LA). These findings confirm that I. galbana IgFAD2 plays important roles in the biosynthetic pathways of unsaturated fatty acids.

关键词： delta-12 fatty acid desaturase expression analysis Isochrysis galbana

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Cloning a Soluble Acyl-ACP desaturase from Marine Microalgae ***

Cloning a Soluble Acyl-ACP Desaturase from Marine Microalgae...

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中国细胞生物学学会第九次会员代表大会暨青年学术大会

作者： Dong-Juan Yuan~1 Ke-Yuan Zhou~1 Jing-Xuan Kang~2 Li-Ming Jiang~1 1 Department of Biochemistry and Molecular Biology,Guangdong Medical College,Zhanjiang 524023,China 2 Department of Medicine,Massachusetts General Hospital and Harvard Medical School,Boston,MA 02114,USA

Background:Marine unicell microalgae had been extensively used as bait in aquaculture because it contains rich health beneficial of protein and unsaturated fatty *** this experiment,after GC-MS was used to detect PUFA contents of six microalgae,several kinds of PCR ways were utility to clone fatty acid desaturase gene from microalgae.

关键词： Acyl-ACP desaturase RLM-RACE PCR Pavlova viridis

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Estimation of the Tissue Distribution of mRNA Transcripts for desaturase and Elongase Enzymes in Channa striata （Bloch, 1793） Fingerlings using PCR Technique

Estimation of the Tissue Distribution of mRNA Transcripts fo...

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2012 4th International Conference on Agriculture and Animal Science

作者： Mohammed Aliyu-Paiko Annette Jaya-Ram Alexander Chong S. C. Roshada Hashim Amuzat A. Olalekan Laboratory of Feeds development and Feeding Management Aquaculture Research Group School of Biological Sciences Universiti Sains Malaysia Laboratory of Fish Biology Aquaculture Research Group School of Biological Sciences Universiti Sains Malaysia

The variations in the capacity of fish species to biosynthesize the n-3 highly polyunsaturated fatty acids （HUFA）, eicosapentaenoic （EPA） and docosahexaenoic （DHA） acids which are crucial to the health of higher vertebrates has been well documented. HUFA biosynthesis has been reported to involve enzyme-mediated fatty acyl desaturation and elongation processes. In previous studies, various authors have cloned, characterized and identified different genes for fatty acyl desaturase and elongase （FAD and FAE, respectively） enzymes from fish species such as Atlantic salmon and zebra fish, and have also shown the up and down regulation of the activity of these enzymes in response to fluctuations in dietary HUFA. In this paper, we report on the tissue distribution in the expression of genes （mRNA） for FAD and FAE enzymes in Channa striata （Bloch, 1793） fingerlings, to evaluate the tissues of the fish which show the highest activity of these enzymes. For this purpose, mRNA from 8 different tissues of the fish （maintained on a commercial diet） were isolated and analyzed, using conventional PCR technique. The tissue distribution of the genes was estimated to be significantly （P＜0.05） higher for both FAD and FAE enzymes in the liver and brain of C. striata than in the other 6 tissues measured （muscle, ovary, testis, gill and skin） respectively. The implication of this observation with respect to the use of vegetable oils to substitute fish oil in diets for freshwater fish species is discussed accordingly.

关键词： Channa striata PCR desaturase Elongase HUFA mRNA expression

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Generation of fad2 transgenic mice that produce omega-6 fatty acids

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Science China(Life Sciences) 2009年第11期52卷 1048-1054页

作者： CHEN Qing1, LIU Qing2, WU ZhiFang1, WANG ZongYi3 & GOU KeMian1 1 State Key Laboratory for Agrobiotechnology, China Agricultural University, Beijing 100193, China 2 Blackmountain Laboratory, CSIRO Plant Industry, Canberra, Australia 3 College of Animal Science and Technology, China Agricultural University, Beijing 100193, China State Key Laboratory for Agrobiotechnology China Agricultural University Beijing China Blackmountain Laboratory CSIRO Plant Industry Canberra Australia College of Animal Science and Technology China Agricultural University Beijing China

Fatty acid desaturase-2 (FAD2) introduces a double bond in position 12 in oleic acid (18:1) to form linoleic acid (18:2 n-6) in higher plants and microbes. A new transgenic expression cassette, containing CMV promoter/fad2 cDNA/SV40 polyA, was constructedto produce transgenic mice. Among 63 healthy offspring, 10 founders (15.9%) integrated the cotton fad2 transgene into their genomes, as demonstrated by PCR and Southern blotting analysis. All founder mice were fertile and heterozygous fad2 female and nontransgenic littermates were used for fatty acid analysis using gas chromatography. One fad2 transgenic line showed substantial differences in the fatty acid profiles and the level of linoleic acid was increased 19% (P<0.05) in transgenic muscles compared to their nontransgenic littermates. Moreover, it exhibited an 87% and a 9% increase (P<0.05) in arachidonic acid (20:4 n-6) in muscles and liver, compared to their nontransgenic littermates. The results indicate that the plant fad2 gene can be functionally expressed in transgenic mice and may playan active role in conversion of oleic acid into linoleic acid.

关键词： fatty acid desaturase FAD2 transgenic mouse oleic acid linoleic acid arachidonic acid

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Effect of Dietary Lipid on the Growth, Fatty Acid Composition and Δ5 Fads Expression of Abalone(Haliotis discus hannai Ino) Hepatopancreas

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Journal of Ocean University of China 2015年第2期14卷 317-324页

作者： LI Mingzhu MAI Kangsen AI Qinghui HE Gen XU Wei ZHANG Wenbing ZHANG Yanjiao ZHOU Huihui LIUFU Zhiguo The Key Laboratory of Aquaculture Nutrition and Feed of Ministry of Agriculture The Key Laboratory of Mariculture of Ministry of Education Ocean University of China College of Agriculture Ludong University

This study investigated the effect of dietary lipid on the growth, fatty acid composition and Δ5 fatty acyl desaturase genes(Fads) expression of juvenile abalone(Haliotis discus hannai Ino) hepatopancreas. Six purified diets were formulated to contain tripalmitin(TP), olive oil(OO, 72.87% 18:1n-9), grape seed oil(GO, 68.67% 18:2n-6), linseed oil(LO, 70.48% 18:3n-3), ARA oil(AO, 41.81% ARA) or EPA oil(EO, 44.09% EPA and 23.67% DAH). No significant difference in survival rate was observed among abalone fed with different diets. Weight gain rate(WGR) and daily growth rate of shell length(DGRSL) were significantly increased in abalone fed with diets containing OO, AO and EO, but decreased in abalone fed with LO diet(P < 0.05) in comparison with those fed with TP. High level of dietary 18:2n-6 resulted in higher content of n-6 polyunsaturated fatty acids(PUFAs) in abalone fed with GO than those fed with TP, OO, LO and EO(P < 0.05). n-3 PUFAs in abalone fed with LO was significantly higher than those in abalone fed with TP, OO, GO and AO(P < 0.05). The highest contents of 20:1n-9 and 22:1n-9 were observed in abalone fed with OO. The expression of Δ5 Fads in hepatopancreas of abalone was enhanced by high concentration of 18:3n-3 and suppressed by dietary LC-PUFAs; however it was not affected by dietary high concentration of 18:1n-9 or 18:2n-6. These results provided valuable information for understanding the synthesis of LC-PUFAs and nutritional regulation of Δ5 Fads expression in abalone.

关键词： desaturase fatty acid gene expression, fish oil vegetable oil

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