Morris S M, Nilson J H, Jenik R A, Winberry L K, McDevitt M A, Goodridge A G
J Biol Chem. 1982 Mar 25;257(6):3225-9.
A double-stranded cDNA library was constructed using total poly(A)+ RNA from the goose uropygial gland. Clones containing sequences complementary to fatty acid synthase mRNA were initially identified by colony hybridization with a 32P-labeled cDNA transcribed from RNA enriched for fatty acid synthase mRNA. Identity of the fatty acid synthase clones was confirmed by hybrid-selected translation. Mature fatty acid synthase mRNA is approximately 16 kilobases in length. When unfed neonatal goslings were fed for 24 hr, relative synthesis of hepatic fatty acid synthase increased more than 42-fold. Concomitantly, hepatic fatty acid synthase mRNA levels increased 70-fold. Thus, nutritional regulation of the synthesis of hepatic fatty acid synthase probably occurs at the pretranslational level. The availability of a specific probe for fatty acid synthase mRNA should allow us to analyze the regulation of expression of this gene during development, by nutrition and by hormones in both liver and uropygial gland.
利用来自鹅尾脂腺的总聚腺苷酸加尾RNA构建了双链cDNA文库。最初通过菌落杂交，用从富含脂肪酸合酶mRNA的RNA转录而来的32P标记cDNA，鉴定出含有与脂肪酸合酶mRNA互补序列的克隆。脂肪酸合酶克隆的身份通过杂交选择翻译得到确认。成熟的脂肪酸合酶mRNA长度约为16千碱基。未喂食的新生雏鹅喂食24小时后，肝脏脂肪酸合酶的相对合成增加了42倍以上。同时，肝脏脂肪酸合酶mRNA水平增加了70倍。因此，肝脏脂肪酸合酶合成的营养调节可能发生在翻译前水平。脂肪酸合酶mRNA特异性探针的可用性将使我们能够分析该基因在发育过程中、在肝脏和尾脂腺中受营养和激素调节的表达情况。
