Molecular cloning of cDNA sequences coding for the major α-and β-globin polypeptides of adult Xenopus laevis

• Published in: Nucleic acids research 1980-06, Vol.8 (12), p.2691-2708
• Main Authors: Kay, R.M., Harris, R., Patient, R.K., Williams, J.G.
• Format: Article
• Language: English
• Subjects: Animals; Base Sequence; Cloning, Molecular; DNA, Recombinant - metabolism; Escherichia coli - metabolism; Female; Globins - biosynthesis; Molecular Weight; Nucleic Acid Conformation; Nucleic Acid Hybridization; Plasmids; Protein Biosynthesis; RNA, Messenger - biosynthesis; Transcription, Genetic; Xenopus; Xenopus laevis
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/8.12.2691

This report describes the synthesis and cloning of almost complete DNA copies of the mRNAs encoding the major α-globin and major β-globin of X.laevis. Double-stranded globin cDNA was inserted into the PstI site of the plasmid pBR322 and two cloned recombinants (designated pXG6C1 and pXG8D2) were selected. These were shown to contain almost complete copies of X.laevis globin mRNA. Restriction enzyme maps were determined for each cDNA sequence using the established method of partial digestion of end labelled DNA. However, this procedure was modified such that isolation of individual DNA fragments was no longer required. Each plasmid was shown, by both hybrid arrested translation and filter selection of complementary RNA, to contain a sequence coding for one or other of the two major globin polypeptides. Sufficient DNA sequence information has been determined from each cDNA clone to demonstrate that pXG8D2 contains β-globin sequence and pXG6C1 contains an α-globin sequence.