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DNA sequence analysis near the Arabidopsis thaliana ABI3 gene revealed the presence of a non-LTR retrotransposon insertion that we have designated Ta11-1. This insertion is 6.2 kb in length and encodes two overlapping reading frames with similarity to non-LTR retrotransposon proteins, including reverse transcriptase. A polymerase chain reaction assay was(More)
The nonrandom integration of retrotransposons and retroviruses suggests that chromatin influences target choice. Targeted integration, in turn, likely affects genome organization. In Saccharomyces, native Ty5 retrotransposons are located near telomeres and the silent mating locus HMR. To determine whether this distribution is a consequence of targeted(More)
The yeast retrotransposon Ty5 preferentially integrates into regions of silent chromatin. Ty5 cDNA also recombines with homologous sequences, generating tandem elements or elements that have exchanged markers between cDNA and substrate. In this study, we demonstrate that Ty5 integration depends upon the conserved DD(35)E domain of integrase and cis-acting(More)
Retroelement cDNA can integrate into the genome using the element-encoded integrase or it can recombine with preexisting elements using the recombination system of the host. Recombination is a particularly important pathway for the yeast retrotransposon Ty5 and accounts for approximately 30% of the putative transposition events when a homologous substrate(More)
Retrotransposons and retroviruses replicate by reverse transcription of an mRNA intermediate. Most retroelements initiate reverse transcription from a host-encoded tRNA primer. DNA synthesis typically extends from the 3'-OH of the acceptor stem, which is complementary to sequences on the retroelement mRNA (the primer binding site, PBS). However, for some(More)
The Saccharomyces retrotransposon Ty5 integrates preferentially into transcriptionally inactive regions (silent chromatin) at the HM loci and telomeres. We found that silent chromatin represses basal Ty5 transcription, indicating that these elements are encompassed by silent chromatin in their native genomic context. Because transcription is a requirement(More)
Permanent records of the results of electrophoretic separations of radiolabeled proteins and membrane-bound proteins (and RNA and DNA) can be made using autoradiography, fluorography, and phosphor imaging. These images can subsequently be quantified using densitometry to obtain a relative measure of the amount of radioactivity in a sample. This unit also(More)
Homologues of Topo IB were retrieved from the nr and the environmental database (including sequences from the GOS project [1]) at the NCBI (http://www.ncbi.nlm.nih.gov/) using the BLASTp software [2]. For environmental sequences, only nearly complete sequences (i.e. longer than 350 amino acids positions) having an e-value lower than 10 using the sequence(More)
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