The effect of gene polymorphism in promoter and intron 1 on human ApoA I expression / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 610-613, 2006.
Artigo
em Chinês
| WPRIM
| ID: wpr-285068
ABSTRACT
<p><b>OBJECTIVE</b>To construct pGL2-ApoA I luciferase reporter vector containing ApoA I gene regulation area, and to investigate the effect of G --> A and C --> T substitution in ApoA I promoter -75 bp and intron 1 +83 bp region respectively on ApoA I gene expression.</p><p><b>METHODS</b>Human chromosome DNA fragments containing ApoA I gene were amplified by PCR, and the DNA fragments consisting of ApoA I AA/CC, GG/TT and GG/CC genetypes were selected separately, then pUC vector including above three different DNA fragments was constructed. After digesting pUC vector with Sac I and Bgl II, ligate the different DNA fragments to basic pGL2 vector that containing luciferase reporter gene. Recombinant and PRL-null vector were cotransfected into HepG2 cells by using cationic liposome method. Cells were cultured for 48 h, activity of firefly and renills luciferase was measured.</p><p><b>RESULTS</b>Three vectors with pGL2-ApoA I-L(-2500 to +289 bp) long fragment vectors and 3 with pGL2-ApoA I-S(-145 to +289 bp) short fragment vectors were combinated successfully. Relative activity of luciferase for ApoA I AA/CC or GG/TT was lower than that for GG/CC significantly.</p><p><b>CONCLUSION</b>-75 bp G --> A and +83 bp C --> T substitution in ApoA I gene may inhibit ApoA I gene transcription and expression. It may be the reason why subjects containing -75 bp A and +83 bp T have lower high density lipoprotein cholesterol (HDL-C) concentration.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas Recombinantes de Fusão
/
Íntrons
/
Transfecção
/
Regiões Promotoras Genéticas
/
Apolipoproteína A-I
/
Mutação Puntual
/
Polimorfismo de Nucleotídeo Único
/
Linhagem Celular Tumoral
/
Vetores Genéticos
/
Genética
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Medical Genetics
Ano de publicação:
2006
Tipo de documento:
Artigo
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