Thiede M A, Ozols J, Strittmatter P
J Biol Chem. 1986 Oct 5;261(28):13230-5.
Hepatic poly(A+) RNA from rats induced for stearyl-CoA desaturase was used for primer-extension of cDNA coding for stearyl-CoA desaturase. Previously, Northern blot analysis showed that translatable desaturase mRNA is 4,900 nucleotides in length (Thiede, M. A., and Strittmatter, P. (1985) J. Biol. Chem. 260, 14459-14463). Six overlapping cDNAs, ranging from 850 to 1450 bases, were used to compile the 4,689-nucleotide sequence. The cDNA includes a 1,074-base open reading frame coding for 358 amino acids, corresponding to a molecular mass of 41,400 daltons. Positive identification of this open reading frame was accomplished by matching the amino acid sequence of both amino-terminal and cyanogen bromide peptides of the purified enzyme with regions of the sequence deduced from the cDNA. Amino acid composition data from the cDNA compares well with that from the desaturase. The protein contains 62% hydrophobic amino acids. An interesting feature of this mRNA is the 3,500-base 3' noncoding region, which has been localized on a single 3' exon by Southern blot analysis.
从经硬脂酰辅酶A去饱和酶诱导的大鼠肝脏中提取的多聚腺苷酸（poly(A+)）RNA，用于对编码硬脂酰辅酶A去饱和酶的cDNA进行引物延伸。此前，Northern印迹分析表明，可翻译的去饱和酶mRNA长度为4900个核苷酸（蒂德，M. A.，和斯特里特马特，P.（1985年）《生物化学杂志》260，14459 - 14463）。使用六个重叠的cDNA（长度从850到1450个碱基）来汇编4689个核苷酸的序列。该cDNA包含一个1074个碱基的开放阅读框，编码358个氨基酸，对应分子量为41400道尔顿。通过将纯化酶的氨基末端和溴化氰肽的氨基酸序列与从cDNA推导的序列区域进行匹配，完成了对这个开放阅读框的阳性鉴定。来自cDNA的氨基酸组成数据与来自去饱和酶的氨基酸组成数据匹配良好。该蛋白质含有62%的疏水氨基酸。这个mRNA的一个有趣特征是其3500个碱基的3'非编码区，通过Southern印迹分析已将其定位在单个3'外显子上。
