title: [Novel bidirectional promoter from human genome].
abstract: In human and other mammalian genomes a number of closely linked gene pairs transcribed in opposite directions are found. According to bioinformatic analysis up to 10% of human genes are arranged in this way. In present work the fragment of human genome was cloned that separates genes localized at 2p13.1 and oriented "head-to-head", coding for hypothetical proteins with unknown functions--CCDC (Coiled Coil Domain Containing) 142 and TTC (TetraTricopeptide repeat Containing) 31. Intergenic CCDC142-TTC31 region overlaps with CpG-island and contains a number of potential binding sites for transcription factors. This fragment functions as bidirectional promoter in the system ofluciferase reporter gene expression upon transfection of human embryonic kidney (HEK293) cells. The vectors containing genes of two fluorescent proteins--green (EGFP) and red (DsRed2) in opposite orientations separated by the fragment of CCDC142-TTC31 intergenic region were constructed. In HEK293 cells transfected with these vectors simultaneous expression of two fluorescent proteins is observed. Truncated versions of intergenic region were obtained and their promoter activity measured. Minimal promoter fragment contains elements Inr, BRE, DPE characteristic for TATA-less promoters. Thus, from the human genome the novel bidirectional promoter was cloned that can be used for simultaneous constitutive expression of two genes in human cells.
authors.forename: A S
authors.initials: AS
authors.lastname: Orekhova
authors.forename: P S
authors.initials: PS
authors.lastname: Sverdlova
authors.forename: P V
authors.initials: PV
authors.lastname: Spirin
authors.forename: O G
authors.initials: OG
authors.lastname: Leonova
authors.forename: V I
authors.initials: VI
authors.lastname: Popenko
authors.forename: V S
authors.initials: VS
authors.lastname: Prasolov
authors.forename: P M
authors.initials: PM
authors.lastname: Rubtsov
mesh_terms: D001483:Base Sequence; D002889:Chromosomes, Human, Pair 2 / Q000235:genetics; D003001:Cloning, Molecular; D018899:CpG Islands; D021901:DNA, Intergenic / Q000235:genetics*; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D015894:Genome, Human / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011506:Proteins / Q000235:genetics*; D017384:Sequence Deletion; D014158:Transcription, Genetic
chemical_list: C560779:CCDC142 protein, human; D021901:DNA, Intergenic; D011506:Proteins; C560780:TTC31 protein, human