caa a22 4500 445318104 CHVBK 20180317142654.0 cr unu---uuuuu 170323e20110201xx s 000 0 eng 10.1007/s10126-010-9262-6 doi (NATIONALLICENCE)springer-10.1007/s10126-010-9262-6 Cloning and Functional Identification of Delta5 Fatty Acid Desaturase Gene and Its 5′-Upstream Region from Marine Fungus Thraustochytrium sp. FJN-10 [Elektronische Daten] [Jian-Zhong Huang, Xian-Zhang Jiang, Xiao-Feng Xia, Ai-Qun Yu, Ruo-Yu Mao, Xiao-Feng Chen, Bao-Yu Tian] A gene encoding delta5 fatty acid desaturase (fad5) was cloned from marine fungus Thraustochytrium sp. FJN-10, a species capable of producing docosahexaenoic acid. The open reading frame of fad5 was 1,320bp and encoded a protein comprising 439 amino acids. Expression of the fad5 in Saccharomyces cerevisiae INVSC1 revealed that FAD5 is able to introduce a double bond at position 5 of the dihomo-γ-linolenic acid (20:3 Δ8,11,14), resulting in arachidonic acid (20:4 Δ5,8,11,14) with a conversion rate of 56.40% which is the highest among engineering yeasts reported so far. The 5′-upstream region of fad5 was cloned by LA-PCR and analyzed. Phylogenetic analysis of this sequence with the 5′-upstream region of other delta5 desaturases showed that the 5′-upstream region of fad5 from Thraustochytrium share the smallest evolution distance with human and rhesus. Computational analysis of the nucleotide sequence of the 5′-upstream region of fad5 has revealed several basic transcriptional elements including five TATA boxes, three CCAAT boxes, 12 GC boxes, and several putative target-binding sites for transcription factors such as HSF, CAP, and ADR1. Preliminary functional analysis of this promoter in S. cerevisiae shows that the 5′-upstream region of fad5 could drive the expression of green fluorescent protein. Springer Science+Business Media, LLC, 2010 Desaturase nationallicence Promoter nationallicence Thraustochytrium nationallicence Polyunsaturated fatty acid nationallicence Docosahexaenoic acid nationallicence Huang Jian-Zhong Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut Jiang Xian-Zhang Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut Xia Xiao-Feng Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut Yu Ai-Qun Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut Mao Ruo-Yu Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut Chen Xiao-Feng Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut Tian Bao-Yu Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut Marine Biotechnology Springer-Verlag 13/1(2011-02-01), 12-21 1436-2228 13:1<12 2011 13 10126 https://doi.org/10.1007/s10126-010-9262-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10126-010-9262-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Huang Jian-Zhong Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut NATIONALLICENCE 700 1- Jiang Xian-Zhang Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut NATIONALLICENCE 700 1- Xia Xiao-Feng Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut NATIONALLICENCE 700 1- Yu Ai-Qun Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut NATIONALLICENCE 700 1- Mao Ruo-Yu Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut NATIONALLICENCE 700 1- Chen Xiao-Feng Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut NATIONALLICENCE 700 1- Tian Bao-Yu Engineering Research Center of Industrial Microbiology, Ministry of Education, Fujian Normal University, 350108, Fuzhou, Fujian, People's Republic of China aut NATIONALLICENCE 773 0- Marine Biotechnology Springer-Verlag 13/1(2011-02-01), 12-21 1436-2228 13:1<12 2011 13 10126 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer