Molecular cloning and organization of two leghaemoglobin genomic sequences of soybean

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Scientific paper

The leghaemoglobins (Lb) are myoglobin-like proteins found in all nitrogen-fixing root nodules of legumes1-3. They are encoded by plant nuclear genes4 which are specifically induced and form the predominant protein in nodules developed in symbiosis with the appropriate species of Rhizobium. The Lb is located in the host-cell cytoplasm of the infected cell5 and is thought to facilitate oxygen diffusion6,7. Amino acid sequencing of the soybean Lbs has revealed at least four primary structures differing only in a few amino acids8-10. We have previously estimated about 40 copies of Lb sequences in the soybean (Glycine max L.) genome by cDNA hybridization4. To investigate Lb gene organization and function, we prepared and characterized a Lb cDNA recombinant molecule, pLb1, and used it to isolate two genomic Lb sequences from a library constructed in Charon 4. We report here that the organization of the two genomic Lb sequences is quite distinct and one of them seems to have an intervening sequence(s). Hybridization of pLb1 with genomic DNA from various tissues showed that Lb sequences are dispersed through more than 30 kilobases of genomic DNA and that there is no apparent sequence rearrangement or methylation changes following induction of Lb genes.

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