DNA sequence organization in soybean investigated by electron microscopy

Abstract

An electron microscopic analysis of the DNA sequence organization in the soybean genome is reported. This analysis employed the gene 32 proteinethidium bromide spreading technique, a procedure which produces striking contrast between double and single-stranded DNA regions. To investigate the arrangement of repetitive sequences differing in genomic frequency, three kinetic fractions of 5 kb DNA fragments were isolated by reassociation and hydroxyapatite chromatography. Renatured structures in each fraction were then visualized in the electron microscope. The majority of repeated sequences, irrespective of frequency, were shown to be relatively non-divergent, to exceed 1.5 kbp in length (number-average), and to be organized primarily into long regularly repeating tandem or clustered arrays. Duplex regions >5 kbp were commonly visualized. A small fraction of low frequency repeats (<100 copies per genome), however, was observed to have a distinctly different form of arrangement. These repeats averaged 0.2 kbp in length, contained divergent sequences, and were contiguous to single copy DNA sequences having an average length of 1.15 kbp. Repeats which flanked a given single copy sequence did not appear to be homologous. Neither short clustered permuted repeats nor interspersion of repeats which differed significantly in reiteration frequency were found to be major features of soybean genome organization.