Construction and sequence of cDNA for rat liver stearyl coenzyme A desaturase

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. C...

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Published inThe Journal of biological chemistry Vol. 261; no. 28; pp. 13230 - 13235
Main Authors Thiede, M A, Ozols, J, Strittmatter, P
Format Journal Article
LanguageEnglish
Published Bethesda, MD Elsevier Inc 05.10.1986
American Society for Biochemistry and Molecular Biology
Subjects
Amino Acid Sequence
Animals
Base Sequence
Biological and medical sciences
Biotechnology
Cyanogen Bromide - pharmacology
DNA - analysis
Fatty Acid Desaturases - genetics
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Liver - enzymology
Methods. Procedures. Technologies
Molecular cloning
Molecular Weight
Poly A - metabolism
Protein Biosynthesis
Rats
RNA - metabolism
RNA, Messenger - metabolism
Stearoyl-CoA Desaturase - analysis
Stearoyl-CoA Desaturase - genetics
Vertebrata
Mammalia
Complementary DNA
Rat
Animal
Liver
Rodentia
Molecular cloning
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Summary: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.
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ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(18)69295-6