Identification and characterization of the repetitive DNA element in the genome of insect iridescent virus type 6

Abstract

The genome of the Chilo iridescent virus (CIV) was analyzed for existence of repetitive DNA sequences by DNA-DNA hybridization using a defined and complete gene library of the viral genome (209 kbp) and by heteroduplex mapping. These experiments revealed the presence of repetitive DNA elements in the CIV genome, which are located in the EcoRI fragment H and in the EcoRI DNA fragment C at the coordinates 0.535 to 0.548 ( EcoRI/ Pstl DNA fragment, 2.7 kbp) and 0.920 to 0.944 ( Pvull CIV DNA fragment L, 5.1 kbp), respectively. The DNA nucleotide sequence (2708 bp) of the EcoRI/ Pstl subfragment was determined. The comparative analysis of the DNA sequences of this particular region of the viral genome with the DNA sequences of the Pvull DNA fragment L (5064 bp) revealed the presence of several DNA sequences within the EcoRI/ Pstl subfragment of the EcoRI CIV DNA fragment H which show homology to DNA sequences of the Pvull DNA fragment L. For example, a DNA element (☐ A, 91 bp) is located at nucleotide positions 1981 to 2072 of the EcoRI CIV DNA fragment H which are complementary (>90%) to the nine regions of the Pvull DNA fragment L (L-☐es 1 to 9). Furthermore heteroduplex mapping revealed the existence of a stem-loop structure (stem, 65 ± 10 by and loop, 652 ± 80 bp) at the genome coordinates 0.571 to 0.582 (2.5 kbp, HindIII/ EcoRI subfragment of the EcoRI CIV DNA fragment H). This indicates that an inverted repeat sequence is located at this region of the viral genome. The DNA nucleotide sequence of this subfragment was determined (2555 bp) which confirmed the data obtained from electron microscopy. An inverted repeat DNA sequence located at nucleotide positions 304 and 1011 is able to form this type of stem-loop structure.