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转座因子Minos的染色体外转座发生在家蚕胚胎中。

Extrachromosomal transposition of the transposable element Minos occurs in embryos of the silkworm Bombyx mori.

作者信息

Shimizu K, Kamba M, Sonobe H, Kanda T, Klinakis A G, Savakis C, Tamura T

机构信息

Department of Biological Science, Konan University, Kobe, Japan.

出版信息

Insect Mol Biol. 2000 Jun;9(3):277-81. doi: 10.1046/j.1365-2583.2000.00182.x.

Abstract

To assess the ability of the transposable element Minos to act as a vector for genetic manipulation of the silkworm Bombyx mori, an extrachromosomal transposition assay based on three plasmids was performed. The three plasmids - helper, donor and target - were co-injected into preblastoderm embryos. Low molecular weight DNA was extracted from the embryos at the stage of blastokinesis and used to transform Escherichia coli. High frequency of transposition was observed in the presence of a helper plasmid possessing an intronless Minos transposase gene, whereas transposition did not occur in the presence of a helper plasmid with the intron-bearing transposase gene. Sequence analysis of the insertion sites showed that Minos always inserts into a TA dinucleotide. Although the insertions are distributed throughout the target gene, there was a preference for certain insertion sites. However, no consensus could be identified in the sequence flanking the target site. The results strongly suggest that the transposable element Minos has the potential to be used as a vector in the silkworm and probably in other lepidopteran insects.

摘要

为了评估转座元件Minos作为家蚕基因操作载体的能力,进行了基于三种质粒的染色体外转座分析。这三种质粒——辅助质粒、供体质粒和靶体质粒——被共同注射到胚盘形成前的胚胎中。在胚动期从胚胎中提取低分子量DNA,并用于转化大肠杆菌。在含有无内含子Minos转座酶基因的辅助质粒存在的情况下观察到高频转座,而在含有带内含子转座酶基因的辅助质粒存在的情况下则不发生转座。插入位点的序列分析表明,Minos总是插入到TA二核苷酸中。虽然插入分布在整个靶基因中,但对某些插入位点有偏好。然而,在靶位点侧翼序列中未发现共有序列。结果强烈表明,转座元件Minos有潜力在家蚕以及可能在其他鳞翅目昆虫中用作载体。

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