We have determined the genomic sequences of four virulent myophages, Bcep1, Bcep43, BcepB1A, and Bcep781, whose hosts are soil isolates of the Burkholderia cepacia complex. Despite temporal and spatial separations between initial isolations, three of the phages (Bcep1, Bcep43, and Bcep781, designated the Bcep781 group) exhibit 87% to 99% sequence identity to one another and most coding region differences are due to synonymous nucleotide substitutions, a hallmark of neutral genetic drift. Phage BcepB1A has a very different genome organization but is clearly a mosaic with respect to many of the genes of the Bcep781 group, as is a defective prophage element in Photorhabdus luminescens. Functions were assigned to 27 out of 71 predicted genes of Bcep1 despite extreme sequence divergence. Using a lambda repressor fusion technique, 10 Bcep781-encoded proteins were identified for their ability to support homotypic interactions. While head and tail morphogenesis genes have retained canonical gene order despite extreme sequence divergence, genes involved in DNA metabolism and host lysis are not organized as in other phages. This unusual genome arrangement may contribute to the ability of the Bcep781-like phages to maintain a unified genomic type. However, the Bcep781 group phages can also engage in lateral gene transfer events with otherwise unrelated phages, a process that contributes to the broader-scale genomic mosaicism prevalent among the tailed phages.

A new type of analytical and preparative cytometric instrument was developed. fie instrument ccxnbines image analysis and machine vision with single cell and chromosome manipulation by means of optical trapping. A proof-of-@nciplc instrument, OCAM, has the ability to locate and analyze biological particles inside an exlosed manipulation chamber, as well as the ability to move and position particIes =card.ing to prcprogmmm ed pocols.Prelimirlfu-y results and potential biologicaJ applications of such a microrobcx are discussed.

The coenzyme A-synthesizing protein complex (CoA-SPC) is a multienzyme complex of Saccharomyces cerevisiae (Bakers' yeast), which has a molecular weight in excess of 200,000 as determined by Sephadex G-200 column chromatography. This multienzyme complex, which is insoluble in the crude yeast cell lysate, has been purified 229-fold. A cellular component of the yeast cell lysate, referred to as t-Factor, with a molecular weight of 400-1000 and chloride ion are involved in the solubilization of CoA-SPC. The CoA-SPC requires L-cysteine, D-pantothenic acid and ATP as substrates. The terminal CoA-SPC-bound intermediate is dephospho-CoA, which is subsequently phosphorylated and released from the complex as CoA. The sequence of reactions for the synthesis of CoA by the CoA-SPC differs significantly from those previously proposed for other systems. It could be that the reaction sequence is unique for the yeast cell.

Summary.-A transplantable reticulum-cell sarcoma induced by Rauscher virus (RV) in (C57BL/6 x DBA/2)F1 (BDF1) mice was grown in tissue culture. Four separate cell lines were established, all of which grew predominantly in suspension. The doubling time of the cells from these cultures ranged from 17 to 32 h. Each culture continued to replicate RV, as indicated by the infectivity in newborn mice of all fluids tested up to the 75th passage. Since the morphological appearance of the cells in vitro was consistent with that of proerythroblasts, all cultures were tested for their ability to differentiate along the erythrocytic line under the influence of dimethylsulphoxide (DMSO). One of the cultures produced small quantities of haemoglobin independently of DMSO. Another was shown to produce haemoglobin, as well as to take up 59Fe and incorporate it into haem, only in the presence of DMSO. The 2 remaining cultures failed to produce haemoglobin, either spontaneously or in the presence of DMSO. Cells from each of the RV-induced cultures, when inoculated back into BDF1 mice, induced typical reticulum-cell sarcomas, without in vivo evidence of erythroid differentiation. In contrast, 2 morphologically identical but non-infectious cell lines derived from Friend virus-induced reticulumcell sarcomas did not show erythroid differentiation in vivo or in vitro, either in the absence or presence of DMSO.

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