Blinov A G, Sobanov Y V, Bogachev S S, Donchenko A P, Filippova M A
Institute of Cytology and Genetics, Siberian Department of the Russian Academy of Sciences, Novosibirsk, Prospect Lavrentjeva.
Mol Gen Genet. 1993 Mar;237(3):412-20. doi: 10.1007/BF00279446.
Nineteen recombinant phages containing DNA from the region of Balbiani ring a (BRa), which develops on chromosome IV in cells of the special lobe of the Chironomus thummi salivary gland, were isolated from a Chironomus thummi genomic library. Three of the clones contained transposable element sequences that hybridized to more than 100 sites on all four Chironomus chromosomes, including constant and variable sites. Two handogous clones, lambda 24 (which lacks the transposable element) and lambda 43 (which contains this insertion) were investigated by nucleotide sequence analysis. The complete nucleotide sequence of the 4.8 kb transposable element from Chironomus thummi (NLR1Cth) is reported here. This element contains two overlapping open reading frames of 1887 (ORF1) and 2649 bp (ORF2). Three cysteine motifs are found in the sequence of ORF1. Sequence similarity was found between ORF2 and known genes of viruses and transposable elements which encode reverse transcriptase. The NLR1Cth element has no long terminal repeats and is flanked by short direct repeats of the sequence TATCACTGACAAC. A 24 bp poly(dA) sequence was found at the 3' end of the element. Based upon its structural organization and comparative analysis of its nucleotide sequence we suggest that this NLR1Cth element belongs to the class of non-LTR retrotransposons. The genomic clone pC6.10 was previously obtained by microdissection and cloning of DNA from polytene chromosome IV of Chironomus thummi. A 2.4 kb insertion contained part of the 3' terminal region of the NLR1Cth element, but this differed in sequence from the first copy by several nucleotide substitutions and a shorter poly (dA) tract at the 3' end.(ABSTRACT TRUNCATED AT 250 WORDS)
从摇蚊基因组文库中分离出19个重组噬菌体,其含有来自巴尔比亚尼环a(BRa)区域的DNA,该区域在摇蚊唾液腺特殊叶细胞的第四条染色体上发育。其中三个克隆含有可转座元件序列,这些序列可与所有四条摇蚊染色体上的100多个位点杂交,包括恒定位点和可变位点。通过核苷酸序列分析研究了两个同源克隆,λ24(不含可转座元件)和λ43(含有此插入片段)。本文报道了来自摇蚊的4.8 kb可转座元件(NLR1Cth)的完整核苷酸序列。该元件包含两个重叠的开放阅读框,分别为1887 bp(ORF1)和2649 bp(ORF2)。在ORF1序列中发现了三个半胱氨酸基序。在ORF2与编码逆转录酶的病毒和可转座元件的已知基因之间发现了序列相似性。NLR1Cth元件没有长末端重复序列,其两侧是序列TATCACTGACAAC的短直接重复序列。在该元件的3'末端发现了一个24 bp的聚(dA)序列。基于其结构组织和核苷酸序列的比较分析,我们认为该NLR1Cth元件属于非LTR逆转录转座子类别。基因组克隆pC6.10先前是通过显微切割和克隆来自摇蚊多线染色体IV的DNA获得的。一个2.4 kb的插入片段包含NLR1Cth元件3'末端区域的一部分,但该序列与第一个拷贝在几个核苷酸取代和3'末端较短的聚(dA)片段上有所不同。(摘要截短至250字)