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Sequence comparison of MLEs of some lepidopteran insects inhabiting in east Asia isolated by PCR method using the inverted terminal repeat of Cecropia MLE and the analysis of transposase activity of MLE isolated from Attacus atlas of Yonaguni island by th

利用天蚕MLE反向末端重复序列对东亚地区部分鳞翅目昆虫MLE进行序列比对及与那国岛Attacus atlas分离MLE的转座酶活性分析

基本信息

批准号：
14560048
负责人：
NAKAJIMA Yumiko
金额：
$ 2.24万
依托单位：
University of the Ryukyus
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

项目摘要

Mariner-like elements(MLEs) were isolated from several Asian insects(mainly lepidopteran insects) and invertebrates using the inverted terminal repeat(ITR) of the Hyalophora cecropia MLE as PCR primers. Around the discrete 1.3 kbp single bands from some specimens expected to contain the full-length of MLEs and various lengths of the fragments from other insects were detected. The full-length of MLEs isolated from the species inhabiting in relatively close regions around Japan were highly similar to each other classifying into the Cecropia MLEs sub family and some of them had the complete ORFs coding for transposase. This seem to indicate that the horizontal transfer of MLEs has taken place even among phylogenetically remote organisms recently and these MLEs might have the ability to transfer horizontally even now. Some of the clearly inactive MLEs making a complex isolated from Bombyx mori (n=28) and B.mandarina(n-27) were situated in other phylogenetic group than the horizontally group of Cecropia subfamily implies that this type of MLEs had inserted into the ancient genome of these species before the genomic segregation of B.mandarina between the Japanese/Korean types(n=27) and the Chinese/Korean types(n=28). One individual from China had a discrete 4.2-kbp band containing a BMCl and LlBm retrotransposon, which is a unique fragment detected in almost all of the silkworm races and strains(the MLE of this unit was named as BmTNML, Nakajima et al.,1999). Thus, the 4.2-kbp unit might be made spontaneously by crossing with B.mori after the segregation of B.mori genome.The transposase region from the full length of mariner sequence isolated from Attacus atlas, which had the complete ORF for transposase, was cloned into the expression vector pENTR11. And the analyses of its transposition activity in the genome of E.coll and in the cultured cell of B.mori are still progressing now.

以透明杯孢Hyalophora cecropia MLE的反向末端重复序列（ITR）为引物，从几种亚洲昆虫（主要是鳞翅目昆虫）和无脊椎动物中分离到了水手样元件（MLEs）。在分离的1.3kbp的单条带周围，从一些标本中检测到预期含有全长MLE的片段和来自其他昆虫的不同长度的片段。从分布在日本周边较近地区的物种中分离到的MLEs全长序列高度相似，属于Cecropia MLEs亚科，其中一些具有完整的转座酶ORF。这似乎表明，MLE的水平转移已经发生，即使在最近的遗传学远程生物，这些MLE可能有能力水平转移，即使是现在。从家蚕（n=28）和B. michina（n = 27）中分离到的一些明显失活的MLEs形成复合体，位于Cecropia亚科水平群以外的其他系统发育群，这意味着这种类型的MLEs在B. michina在日本/朝鲜型（n=27）和中国/朝鲜型（n=28）之间的基因组分离之前就已经插入到这些物种的古老基因组中。来自中国的一个个体具有含有BMC 1和LlBm反转录转座子的离散的4.2-kbp条带，其是在几乎所有家蚕品种和品系中检测到的独特片段（该单位的MLE被命名为BmTNML，Nakajima et al.，1999年）。因此，4.2 kbp的转座酶片段可能是在分离了B.mori基因组后，通过与B.mori杂交自发产生的。其在大肠杆菌科尔基因组和家蚕B培养细胞中的转座活性分析仍在进行中。