The Incidence of β-thalassemia Heterozygote Car rying α-thalassemia-1 Gene / 中山大学学报(医学科学版)

【Objective】 To investigate the incidence of β-th alassemia (β-thal) heterozygote carrying α-thalassemia (α-thal ) 1 gene in Guangdong area. 【Methods】 β-thal genes were screened by reve rse dot blotting (RDB). In β-thal DNA samples α-thal 1 genes were am plified using gap-PCR method. 【Results】43 α-thal-1 cases were identifi ed among the 500 β-thal traits. The rate is 8.6%. 【Conclusion】 In Guangd ong area the incidence of β-thal heterozygote carrying α-thal-1 gen e is 8.6%, which should be paid much attention in the genetic counselling and pr enatal diagnosis.

Cloning and Sequencing of the Human Thalassemic Gene β654 / 中山大学学报(医学科学版)

【Objective】In order to establish the foundation for transgenic mouse model,the human thalassemic gene(β654) was cloned and sequenced.【Methods】The human β654 gene was amplified by PCR,and cloned into the plasmid BGT51 in which the human β gene was cut out aforehand.The recombinant plasmid was certified by enzyme-digestion,reverse dot hybridization and sequencing.【Results】A recombinant plasmid was obtained,which contained the human β654 gene in the correct recombinant direction.Sequencing showed that the cloned insert was correct.【Conclusions】The recombinant plasmid constructed is useful for establishing a transgenic mouse.

Cloning of human ? thalassemic mutation ? IVS II654(C→T) and establishment of its mammalian expression system / 中国病理生理杂志

AIM: To clone human ?-globin gene carrying a thalassemic mutation IVS II654(C→T) and establish a eukaryotic expression system for high-level expression of human ? IVS II654 gene in mouse erythroleukaemia(MEL) cells. METHODS: The fragments of human ? 654 gene isolated from the ? thalassemia patients homozygous for the ? 654 mutation were amplified by PCR, and cloned to plasmid pBGT51. Then, the human ? LCR and ? 654 gene were subcloned from plasmid pBGT51 to the stable mammalian expression vector pcDNA3.1+ together, and the MEL cells were transfected with this vector using commercially available cationic lipid FuGENE6. The MEL cells were induced for further maturation by DMSO and the expression of human ? 654 gene in the MEL cells was identified by RT-PCR. RESULTS: A mammalian expression system of human ? thalassemic mutation ?IVS II654(C→T) was established. CONCLUSION: The level and the reliability of expression of human ? 654 gene in the MEL cells in vitro are similar to that in vivo in human body. This may be a valuable gene therapy model for human ? thalassemic mutation ?IVS II654(C→T).