Making a construct containing of γ-globin gene and control regions of beta gene cluster aiming for studying of effective mutations in HPFH phenotype
Subject Areas : Journal of Animal Biologyمریم بی خوف تربتی 1 , فاطمه جمشیدی 2 , حسین خان احمد 3 , سیروس زینلی 4 , مرتضی کریمی پور 5
1 - استادیار، گروه زیست شناسی، دانشگاه آزاد اسلامی واحد شهرری
2 - کارشناس ارشد، گروه پزشکی مولکولی، انستیتو پاستورتهران
3 - استادیار ،گروه ب ث ژ، انستیتو پاستورکرج
4 - دانشیار، گروه پزشکی مولکولی، انستیتو پاستورتهران
5 - استادیار، گروه پزشکی مولکولی، انستیتو پاستورتهران
Keywords: andbeta, -thalassemia, Gamma Globin, Fetal hemoglobin, HPFH, LCR,
Abstract :
In andbeta;-thalassemia patients are mutations or deletions in beta globin gene which due to absence or reduction of andbeta;-globin chain synthesis. Adult hemoglobin (HbA) contains 2 Alfa Globin and 2 Beta Globin chains (andalpha;2andbeta;2) while Fetal hemoglobin (HbF) contains 2 Alfa Globin and 2 Gamma Globin chains (andalpha;2andgamma;2). HbF switches to HbA in early weeks after the birth. A kind of andbeta;-thalassemia is HPFH phynotype. In these patients the level of HbF will be high in after the birth continuously. The gamma globin gene is in beta globin gene cluster and its expression will be control by locus control region (LCR) which located in upstream of beta gene cluster. The LCR composed of five DNase I hyper sensitive sites (HS1-5) in Erythroid cells. We amplified HS2, HS3, HS4 segments (mini LCR) and andgamma;-globin gene from the genomic DNA of a normal individual by PCR. Each of segments cloned in pTZ57R/T vector and then subcloned in the pBGGT vector, a derivative of PUC19 plasmid. This construct is a positive control and containing 5and#39; HS4,HS3,HS2 3and#39; segments (mini LCR, 4 kb) and A Gamma gene (2 kb). That could be used in studying of some mutations which due to HPFH phenotype and affect expression of andgamma;-globin gene and also for induction of HbF production in gene therapy of andbeta;-thalassemia.
