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http://dx.doi.org/10.5352/JLS.2004.14.2.315

Expression of Human Serine Palmitoyltransferase Genes for Antibody Development  

김희숙 (경성대학교 식품공학과)
Publication Information
Journal of Life Science / v.14, no.2, 2004 , pp. 315-319 More about this Journal
Abstract
For antibody development of human serine palmitoyltransferase (SPT, EC 2.3.1.50), SPTLC1 and SPTLC2 genes were subcloned in pRset vector and expressed in E. coli BL21 (DE3)pLys cells. Eucaryotic SPT is a membrane-bound heterodimer enzyme, while all other members are soluble homodimer enzymes. cDNA library were obtained from total RNA from human embryo kidney cell line, HEK293, using RT-PCR and PCR with specific primers was carried out for preparing SPTLC1 and SPTLC2 genes. pRset vector which can express hexahistidine-tag fusion protein was used and the DNA sequences of pRsetB/SPTLC1 and pRsetA/SPTLC2 were confirmed. Recombinant BL21 cells with SPTLC subunits were selected with LB plate containing ampicillin and chroramphenicol. SPTLC1 and SPTLC2 proteins were induced with 1 mM IPTG and seperated on 10% SDS-PAGE gel. Expressed proteins were confirmed by western blotting with His-tag antibody.
Keywords
Serine palmitoyltransferase; antibody development; expression;
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  • Reference
1 Mutations in the yeast LCB1 and LCB2 gene, including those corresponding to the hereditary sensory neuropathy type Ⅰ mutations, dominantly inactivate serine palmitoyltransferase /
[ Gable,K.;G.Han;E.Monaghan;D.Bacikova;M.Natarajan;R.Williams;T.M.Dunn ] / J. Biol. Chem.   DOI   ScienceOn
2 Sphingolipid metabolism in the regulation of bioactive molecules /
[ Luberto,C.;Y.A.Hannun ] / Lipids   DOI
3 The biosynthesis of dihydrosphingosine in cell-free preparation of Hansenula ciferri /
[ Braun,P.E.;E.E.Snell ] / Proc. Natl. Acad. Sci. USA   DOI   ScienceOn
4 Human and murine serine-palmitoyl-CoA transferase - cloning, expression and characterization of the key enzyme in sphingolipid synthesis /
[ Weiss,B.;W.Stoffel ] / Eur. J. Biochem.   DOI   ScienceOn
5 Hereditary sensory neuropathy type Ⅰ mutation confer dominant-negative effects on serine palmitoyltransferase, critical for sphingolipid synthesis /
[ Bajaoui,K.;Y.Uchida;S.Yasuda;M.Ho;Nishijima,R.;H.Brown,Jr.;W.M.Holleran;K.Hanada ] / J. Clin. Invest.   DOI
6 Colonic cell proliferation and aberrant crypt foci formation are inhibited by dairy glycophin-golipids in 1,2-dimethylhydrazine-treated CF1 mice /
[ Schmelz,E.M.;M.C.Sullard;D.L.Dillahay;A.H.Merrill,Jr. ] / J. Nutr.
7 Bifuntional role of glycosphingolipids : modulators for transmembrane signaling and mediators for cellular interactions /
[ Hakomori,S. ] / J. Biol. Chem.
8 Ceramide in the eukaryotic stress response /
[ Hannun,Y.A.;C.Luberto ] / Trends Cell Biol.   DOI   ScienceOn
9 Synthesis of C18- and C20- dihydrosphingosines, ketodihydrosphingosines and ceramides by microsomal preparation from mouse brain /
[ Braun,P.E.;P.Morell;N.S.Radin ] / J. Biol. Chem.
10 /
[ Bell,R.M.;Y.A.Hannun;A.H.Merrill,Jr ] / Advances in Lipid Research, Sphingolipid and their metabolite
11 Chemistry of glycosphingolipids /
[ Hakomori,S.;J.N.Kanfer(ed.);S.Hakomori(ed.) ] / Sphingolipid Biochemistry
12 Tsc3p is an 80-amino acid protein associated with serine palmitoyltransferase and required for optimal enzyme activity /
[ Gable,K.;D.Slife;D.Bacikova;E.Monaghan;T.M.Dunn ] / J. Biol. Chem.   DOI   ScienceOn
13 Sphingolipid - Their met-abolic pathway and the pathobiochemistry of neurode-generative disease /
[ Kolter,T.;K.Sandhoff ] / Angew. Chem. Int. Ed.   DOI
14 Two new mutations in the acid sphingomylinase gene causing type a Niemann-pick disease: N389T and R441X /
[ Schuchman,E.H. ] / Hum. Mutat.   DOI   ScienceOn
15 Sphingolipids - the enigmatic lipid class : Biochemistry, physiology and pathophysiology /
[ Merrill,A.H.Jr.;E.M.Schmelz;D.L.Dilehay;S.Spiegel;J.A.Shayman;J.J.Schroeder;R.T.Riley;K.A.Voss;E.Wang ] / Toxicol. Appl. Pharmacol.   DOI   ScienceOn
16 SPTLC1 is mutated in hereditary sensory neuropathy, type Ⅰ /
[ Bejaoui,K.;C.Wu;M.D.Scheffler;G.Hann;P.Ashby;L.Wu;P. de Jong;R.H.Brown,Jr. ] / Nat. Genet   DOI   ScienceOn
17 Signal transduction of stress via ceramide /
[ Mathias,S.;LA.Pena;R.N.Kolesnick ] / Biochem. J.
18 LCB2 gene of Saccharomyces and the related LC81 gene encode subunits of serine palmitoryltransferase, the initial enzyme in sphinglipid synthesis /
[ Nagiec,M.M.;J.A.Baltisberer;G.B.Wells;R.I.Lester;R.C.Dickson ] / Proc. Natl. Acad. Sci. USA   DOI   ScienceOn
19 Sphingolipid metabolism and cell growth regulation /
[ Spiegel,S.;A.H.Merrill,Jr. ] / FASEB J
20 Mutation in SPTLC1, encoding serine palmitoyltransferase, long chain base sub-unit-1, cause hereditary sensory neuropathy type Ⅰ /
[ Dawkins,J.L;D.J.Hulme;S.B.Brahmbhatt;M.Auergrumbach;G.A.Nicholson ] / Nat. Genet.   DOI   ScienceOn
21 A water-soluble homodimeric serine palmitoyltransferase from Sphingomonas paucimobilis EY2395T /
[ Ikushiro,H.;H.Hayashi;H.Kagamiyama ] / J. Biol. Chem.   DOI   ScienceOn
22 Skin barrier structure and function : the single gel phase model /
[ Norlen,L. ] / J. Invest. Dematol.   DOI   ScienceOn
23 Localization, topology and function of the LCB1 subunit of serine palmitoyltransferase in mammalian cells /
[ Yasuda,S.;M.Nishijima;K.Hanada ] / J. Biol. Chem.   DOI   ScienceOn
24 Role of dietary sphing-olipids and inhibitors of sphingolipid metabolism in cancer and other diseases /
[ Merrill,A.H.Jr.;E.M.Schmelz;E.W.Schroeder;D.L.Dillehay;R.T.Reley ] / J. Nutr.