1 The abbreviations used are: KDO, 3-Deoxy-D-manno-octulosonate; KDO 8-P, 3-DeoxyD-manno-octulosonate 8-phosphate; Tris, Tris-(hydroxymethyl)aminomethane; HEPES, N-[2Hydroxyethyl]piperazine-N’-[2-ethanesulfonic acid]; PNPase, purine nucleoside phosphorylase; EDTA, (ethylenedinitrilo)tetraacetic acid; 2D gel electrophoresis, two dimensional gel electrophoresis; SDS-PAGE, sodium dodecyl sulfate-p...

We have used electron microscopy of thin sections and experiments on isolated viroplasms to compare the properties of four strains of cauliflower mosaic virus (CaMV), three of which were partially or completely deleted in open reading frame (ORF) II. Our results confirm that this gene is required for aphid transmissibility and show that the product of ORF II influences the firmness with which v...

We have cloned, sequenced and analyzed transcription of the phage T4 uvsY gene. This gene is transcribed from a single gp MotA-dependent middle promoter to give a major transcript of approximately 930 nucleotides and a minor transcript of approximately 620 nucleotides. All in vivo and in vitro uvsY transcripts show anomalous migration in agarose gels. The uvsY transcript contains an open readin...

The postulated posttranslational modification of the polyhydroxybutyrate (PHA) synthase from Ralstonia eutropha by 4-phosphopantetheine was investigated. Four beta-alanine auxotrophic Tn5-induced mutants of R. eutropha HF39 were isolated, and two insertions were mapped in an open reading frame with strong similarity to the panD gene from Escherichia coli, encoding L-aspartate-1-decarboxylase (E...

For the identification of yeast genes specifying biochemical activities, a genomic strategy that is rapid, sensitive, and widely applicable was developed with an array of 6144 individual yeast strains, each containing a different yeast open reading frame (ORF) fused to glutathione S-transferase (GST). For the identification of ORF-associated activities, strains were grown in defined pools, and ...

RNA interference (RNAi) is a conserved mechanism in which double-stranded, small interfering RNAs (siRNAs) trigger a sequence-specific gene-silencing process. Here we describe the inhibition of murine herpesvirus 68 replication by siRNAs targeted to sequences encoding Rta, an immediate-early protein known as an initiator of the lytic viral gene expression program, and open reading frame 45 (ORF...

The mRNA encoding Escherichia coli polypeptide chain release factor 2 (RF2) has two partially overlapping reading frames. Synthesis of RF2 involves ribosomes shifting to the +1 reading frame at the end of the first open reading frame (ORF). Frameshifting serves an autoregulatory function. The RF2 gene sequences from the 86 additional bacterial species now available have been analyzed. Thirty pe...

Substrate identification is the key to defining molecular pathways or cellular processes regulated by proteases. Although phage display with random peptide libraries has been used to analyze substrate specificity of proteases, it is difficult to deduce endogenous substrates from mapped peptide motifs. Phage display with conventional cDNA libraries identifies high percentage of non-open reading ...

SpoU is encoded by the fourth open reading frame (ORF) of the spo operon of Escherichia coli (1). The 5'-terminal gene of this operon, gmk, encodes guanylate kinase; the second gene, rpoZ, encodes the w subunit of RNA polymerase; the third gene, spoT, encodes ppGpp hydrolase, and the 3'-terminal gene, recG, encodes a helicase-like protein involved in recombination (1 and references therein). Sp...