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Biochemical and electron microscope analyses of the DNA reverse transcripts present in the virus-like particles of the yeast transposon Ty1. Identification of a second origin of Ty1DNA plus strand synthesis.

机译:酵母转座子Ty1病毒样颗粒中存在的DNA反转录产物的生化和电子显微镜分析。 Ty1DNA的第二个起源的鉴定加上链合成。

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摘要

Transposition of Saccharomyces cerevisiae Ty1 retroelements has been shown to involve reverse transcription in intracytoplasmic virus-like particles (Ty-VLPs). Ty DNA present in the particles specified by Ty1-H3 element was found to consist of the full-length genomic DNA as well as incomplete cDNAs mainly of plus polarity. Our results indicate that identical sequences (TGGGTGGTA) are used as primers for the synthesis of plus strand cDNA, generating cDNAs of 0.345 kb (analogous to the retroviral strong-stop plus cDNA) and of 2.1 kb. Electron microscopic analyses of Ty1-VLP DNA revealed two distinct classes, one full-length and the other corresponding to 0.34 kbp molecules, the size of a LTR sequence. The full-length molecules are either completely double-stranded or only partially double- stranded at one end or at both ends. These double-stranded regions are of a length corresponding to those of incomplete plus strands detected by biochemical techniques. Double-stranded circular molecules mainly of a length corresponding to that of two-LTR circles were also detected on electron micrographs. These analyses allowed us to propose a scheme for reverse transcription in Ty particles.
机译:已显示酿酒酵母Ty1逆转录元件的转座涉及胞浆内病毒样颗粒(Ty-VLPs)的逆转录。发现存在于由Ty1-H3元素指定的粒子中的Ty DNA由全长基因组DNA和不完整的cDNA(主要为正极性)组成。我们的结果表明,相同的序列(TGGGTGGTA)被用作合成正链cDNA的引物,产生0.345 kb(类似于逆转录病毒强终止加号cDNA)和2.1 kb的cDNA。 Ty1-VLP DNA的电子显微镜分析揭示了两种不同的类别,一种为全长,另一种对应于0.34 kbp的分子,即LTR序列的大小。全长分子在一端或两端是完全双链的或仅是部分双链的。这些双链区域的长度对应于通过生化技术检测到的不完全加链的长度。在电子显微照片上还检测到主要长度对应于两个LTR环的长度的双链环状分子。这些分析使我们能够提出一种在Ty颗粒中进行反转录的方案。

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