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Nucleic Acids Research, 1981, Vol. 9, No. 17 4391-4402
© 1981

MOLECULAR BIOLOGY

Structure and expression of a cloned ß°thalassaemic globin gene

N. Moschonas, E. de Boer, F.G. Grosveld, H.H.M. Dahl, S. Wright, C.K. Shewmaker and R.A. Flavell

Laboratory of Gene Structure and Expression, National Institute for Medical Research, Mill Hill London NW7 1AA, UK

Received July 2, 1981. We have cloned the single ß-globin gene from an Italian patient who is a double heterozygote for (ß°/{delta}ß° 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 ß-globin. S1-mapping transcription studies show that ß-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-ß°-globin gene recombinants have been transfected into Hela cells and analysed for ß-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 ß-globin mRNA is seen for the ß° thalassaemic globin gene and for a normal ß-globin gene.

To elucidate the nature of the lesion, the entire DNA sequence of the ß-globin gene of this patient has been determined. The sequence shows that this gene contains a termination codon at position 39 (CAG -> UAG). Otherwise, there is a remarkable conservation of the entire DNA sequence.


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