Hitz W D, Carlson T J, Booth J R, Kinney A J, Stecca K L, Yadav N S
E.I. duPont de Nemours & Co., Experimental Station, Wilmington, Delaware 19880-0402.
Plant Physiol. 1994 Jun;105(2):635-41. doi: 10.1104/pp.105.2.635.
Oligomers based on amino acids conserved between known plant omega-3 and cyanobacterium omega-6 fatty acid desaturases were used to screen an Arabidopsis cDNA library for related sequences. An identified clone encoding a novel desaturase-like polypeptide was used to isolate its homologs from Glycine max and Brassica napus. The plant deduced amino acid sequences showed less than 27% similarity to known plant omega-6 and omega-3 desaturases but more than 48% similarity to cyanobacterial omega-6 desaturase, and they contained putative plastid transit sequences. Thus, we deduce that the plant cDNAs encode the plastid omega-6 desaturase. The identity was supported by expression of the B. napus cDNA in cyanobacterium. Synechococcus transformed with a chimeric gene that contains a prokaryotic promoter fused to the rapeseed cDNA encoding all but the first 73 amino acids partially converted its oleic acid fatty acid to linoleic acid, and the 16:1(9c) fatty acid was converted primarily to 16:2(9c, 12) in vivo. Thus, the plant omega-6 desaturase, which utilizes 16:1(7c) in plants, can utilize 16:1(9c) in the cyanobacterium. The plastid and cytosolic homologs of plant omega-6 desaturases are much more distantly related than those of omega-3 desaturases.
基于已知植物ω-3脂肪酸去饱和酶和蓝细菌ω-6脂肪酸去饱和酶之间保守氨基酸的寡聚体,用于筛选拟南芥cDNA文库中的相关序列。一个鉴定出的编码新型去饱和酶样多肽的克隆被用于从大豆和甘蓝型油菜中分离其同源物。植物推导的氨基酸序列与已知植物ω-6和ω-3去饱和酶的相似度低于27%,但与蓝细菌ω-6去饱和酶的相似度超过48%,并且它们含有假定的质体转运序列。因此,我们推断植物cDNA编码质体ω-6去饱和酶。甘蓝型油菜cDNA在蓝细菌中的表达支持了这一鉴定。用一个嵌合基因转化的集胞藻,该嵌合基因包含一个与编码除前73个氨基酸外的油菜籽cDNA融合的原核启动子,在体内将其油酸脂肪酸部分转化为亚油酸,并且16:1(9c)脂肪酸主要转化为16:2(9c, 12)。因此,植物中利用16:1(7c)的植物ω-6去饱和酶在蓝细菌中可以利用16:1(9c)。植物ω-6去饱和酶的质体和胞质同源物之间的亲缘关系比ω-3去饱和酶的同源物更远。