Structure and expression of a cloned β o thalassaemic globin gene.
We have cloned the single beta-globin gene from an Italian patient who is a double heterozygote for beta o/delta beta o thalassaemia. RNA isolated from nucleated red cells from this patient can be translated in vitro to give detectable levels of A gamma- G gamma and alpha-globin, but no beta-globin. S1-mapping transcription studies show that beta-globin mRNA is present at about 1-3% of the level of alpha- and gamma-globin mRNA. In addition, the expression of this gene has been studied by reversed genetics. SV40-plasmid-beta o-globin gene recombinants have been transfected into Hela cells and analysed for beta-globin mRNA. In contrast to the results obtained with mRNA isolated directly from the blood of this patient, in the transfected Hela cells the same level of beta-globin mRNA is seen for the beta o thalassaemic globin gene and for a normal beta-globin gene. To elucidate the nature of the lesion, the entire DNA sequence of the beta-globin gene of this patient has been determined. The sequence shows that this gene contains a termination codon at position 39 (CAG - greater than UAG). Otherwise, there is a remarkable conservation of the entire DNA sequence.
|Keywords||0 (RNA, Messenger), 9004-22-2 (Globins), 9007-49-2 (dna), Base Sequence, Cloning, Molecular, DNA/analysis, Gene Expression Regulation, Globins/genetics, Human, RNA, Messenger/analysis, Simian virus 40, Support, Non-U.S. Gov't, Thalassemia/genetics|
Moschonas, N., Grosveld, F.G., Dahl, H.H.M., Wright, S., Shewmaker, C., Flavell, R.A., & de Boer, E.. (1981). Structure and expression of a cloned β o thalassaemic globin gene.. Nucleic Acids Research, 9(17), 4391–4401. Retrieved from http://hdl.handle.net/1765/2347