title: Molecular cloning, tissue expression and protein structure prediction of the porcine 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGR) gene.
abstract: 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGR) is the rate-limiting enzyme for cholesterol synthesis. Its activity is regulated via a negative feedback mechanism through sterols and non-sterol metabolites derived from mevalonate, the product of the reaction catalyzed by reductase. Here, we cloned a full-length transcript of porcine HMGR by RT-PCR and RACE. The porcine HMGR cDNA (2864 bp,GenBank accession no. DQ432054) contains a 2658 bp open reading frame and shares 91% identify with those of human and cattle, and 85%, 85% and 84% identify with the HMGR coding sequences of Norway rat, golden hamster, and house mouse, respectively. The deduced porcine HMGR protein consists of 885 amino acids with a calculated molecular mass of 97.15 kDa(GenBank accession no. ABD96089). The amino acid sequence similarities correspond to 95%, 95%, 92%, 92% and 92% when compared with human, cattle, Norway rat, golden hamster and house mouse sequences, respectively. The structure and function of HMGR deduced protein product were predicted by bioinformatic approaches. HMGR-specific transcripts were found in 15 different tissues from pig by RT-PCR and Real-time PCR. The relative expression level of HMGR was high in liver, heart, kidney, bladder and subcutaneous fat, medium in lung, uterus and large intestine, and low in cerebrum, spleen, spinal cord, stomach, ovary, longissimus muscle, and small intestine. The SNPs analysis of HMGR showed that there were five SNPS and three of them are synonymous mutations and the other two are missense mutations. Taken together, our data may lay a ground for further investigation of HMGR's functions and regulatory mechanisms in swine.
authors.affiliation: College of Animal Sciences and Technology, National Engineering Laboratory for Animal Breeding & Key Laboratory of Animal Genetics, Breeding and Reproduction, Ministry of Agriculture, P. R. China, China Agricultural University, Beijing 100193, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Chen
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Wang
authors.forename: Zongjin
authors.initials: Z
authors.lastname: Li
authors.forename: Lujun
authors.initials: L
authors.lastname: Kong
authors.forename: Guifen
authors.initials: G
authors.lastname: Liu
authors.forename: Jinluan
authors.initials: J
authors.lastname: Fu
authors.forename: Aiguo
authors.initials: A
authors.lastname: Wang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D025782:Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008969:Molecular Sequence Data; D009154:Mutation; D016366:Open Reading Frames; D020641:Polymorphism, Single Nucleotide; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D013552:Swine / Q000235:genetics*; D001743:Urinary Bladder / Q000201:enzymology; D014599:Uterus / Q000201:enzymology
chemical_list: D025782:Hydroxymethylglutaryl-CoA-Reductases, NADP-dependent