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KMID : 1041220000420010014
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Journal of Sericultural and Entomological Science
2000 Volume.42 No. 1 p.14 ~ p.23
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Sequence Analysis, Molecular Cloning and Restriction Mapping of Mitochondrial Genome of Domesticated Silkworm, Bombyx mori
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Abstract
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The mitochondrial genome of domesticated silkworm (Bombyz mort) was mapped with five restriction endonucleases (BamHI, EcoRI, HrndIII, PstI and Xbal), the entire genome was cloned with HindIII and EcoRI. From the end sequencing results of 5` and 3` region for full genoe set of eleven mitochondrial clones, the seven mitochondrial genes(NADH dehydrogenase 6, ATPase 6, ATPase 8, tRNALys, tRNAASP. tRNAThr and tRNAPhe ) of mori were identified on the basis of their nucleotide sequence homology. The nucleotide composition of NADH dehydrogenase 6 was heavily biased towards adenine and thymine, which accounted for 87.76%. On basis of the sequence similarity with published tRNA genes from six insect species, the tRNALys, tRNAASP. tRNAThr were showed stable canonical clover-leaf tRNA structures with acceptable anticodons. However, both the DHU and TΨC arms of tRNAPhe could not form any stable stem-loop structure. The two overlapping gene paris (tRNALys-tRNAASP and ATPase8-ATPase6) were foud from our sequencing results. The genes are encoded on the same strand. ATPase8 and ATPase6 overlaps(ATGATAA) which are a single example of overlapping events between abutted protein-coding genes are common. and there is evidence that ATPase8 and at an internal start site for ATPase6. Ultimately, this result will provide assistance in designing oligo-nucleotides for PCR amplification, and sequencing the specific mitochondrial genes for phylogenetics of geographic races, genetically improved silkworm strains and wild silkworm (mankarina) which is estimated as ancestral of domesticated silkworm.
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