Hirochika H, Fukuchi A, Kikuchi F
Department of Molecular Biology, National Institute of Agrobiological Resources, Ibaraki, Japan.
Mol Gen Genet. 1992 May;233(1-2):209-16. doi: 10.1007/BF00587581.
Three families of retrotransposons of rice (Tos1, Tos2, and Tos3) were isolated by using a method based on the sequence conservation of the primer binding site for reverse transcription. This method should be generally applicable for cloning retrotransposon of other plants. One retrotransposon, Tos3-1, was studied in detail. Tos3-1 is 5.2 kb long, has structures common to retrotransposons, such as long terminal repeats (LTR), a primer binding site complementary to the initiator tRNA, a polypurine tract, and generates target sequence duplications flanking the inserted element. Southern blotting analysis showed that sequences homologous to Tos1, 2 and 3 are found in wild rice species as well as in cultivated rice species, but not in maize and tobacco. The copy number and genomic location of the families vary in different strains of one species of wild rice, suggesting that these elements may still be active. Retrotransposons were also screened for by amplification of the reverse transcriptase coding region using the polymerase chain reaction (PCR). At least two types of copia-like elements (Tos4 and Tos5) were found. The total copy number of retrotransposons in the rice genome was estimated to be about 1000. These results suggest that, as in Drosophila, retrotransposons are the major transposon class in rice.
通过使用一种基于逆转录引物结合位点序列保守性的方法,分离出了水稻的三类逆转座子(Tos1、Tos2和Tos3)。该方法应该普遍适用于克隆其他植物的逆转座子。对其中一个逆转座子Tos3 - 1进行了详细研究。Tos3 - 1长5.2 kb,具有逆转座子常见的结构,如长末端重复序列(LTR)、与起始tRNA互补的引物结合位点、多聚嘌呤序列,并在插入元件两侧产生靶序列重复。Southern杂交分析表明,在野生稻种和栽培稻种中都发现了与Tos1、2和3同源的序列,但在玉米和烟草中未发现。在一种野生稻的不同菌株中,这些家族的拷贝数和基因组位置各不相同,这表明这些元件可能仍然具有活性。还通过使用聚合酶链反应(PCR)扩增逆转录酶编码区来筛选逆转座子。发现了至少两种类copia元件(Tos4和Tos5)。估计水稻基因组中逆转座子的总拷贝数约为1000。这些结果表明,与果蝇一样,逆转座子是水稻中主要的转座子类。