The state of integration of avian sarcoma virus DNA in the genomes of transformed chicken, duck, and quail fibroblasts was deduced by means of restriction enzyme digestion of total cell DNA, gel electrophoresis, and subsequent analysis by the procedure of Southern. The cells used in these studies were either mass-infected cultures or clones of infected cells selected by their ability to form colonies in agar. For both mass-infected cultures and clones of cells of all three species, we found that integration occurred at a specific site on the viral genome but appeared to occur at many sites on the cell genome. At least some of the integrated viral DNA existed as intact nonpermuted species flanked by direct terminal repeats of at least 0.134 megadalton (217 base pairs). For each of 12 transformed quail clones studied, it was possible to detect, after digestion with Kpn I, unique junctions between viral and cellular DNA. That is, at our level of analysis, the integration site on the cell genome for each clone was different. However, within each of the 17 chicken and 9 duck clones of transformed cells, a heterogeneity presumably occurred during the outgrowth of the cell clone population, in that we could not readily detect identifiable cell-virus junction fragments.

Analysis of integrated avian RNA tumor virus DNA in transformed chicken, duck and quail fibroblasts.

The nucleotidyltransferase encoded by plasmid pUB110 was purified to greater than 95% purity with a 33% yield. The enzyme is a monomeric protein with a molecular weight of 34,000. The optimum pH for activity is 5, and the optimum MgCl2 concentration for activity is 18 mM. The enzyme, which is synthesized constitutively, is stable for several weeks at 4 degrees C. This enzyme would appear to be a good model gene product for the development of a pUB110 deoxyribonucleic acid-dependent in vitro protein-synthesizing system from Bacillus subtilis.

Purification and characterization of a kanamycin nucleotidyltransferase from plasmid pUB110-carrying cells of Bacillus subtilis.

Publisher Summary This chapter has considered sporulation mainly as a problem in the regulation of a complex sequence of events Until now, progress toward a solution of this problem has been slow As a result of genetical analysis to know the dimensions of the problem, progress has been made on the biochemical side The chapter explores that the latest techniques in genetic manipulation have opened up the possibility of cloning sporulation genes and identifying the products for which they code It is now reasonable to hope that within the next few years at least some of the molecules should be identified that govern expression of the sequence and it might be understood, at least in principle, the ways in which they do so Even a partial elucidation of the mechanism should greatly increase the understanding not only of sporulation but of biochemical differentiation in more complex organisms

Early events during bacterial endospore formation.

The structure and mechanism of action of bacterial DNA-dependent RNA polymerase

In vitro studies demonstrated that the Bacillus subtilis subtilisin gene (aprE) could be transcribed by RNA polymerase holoenzyme reconstituted from core and sigma A factor obtained from vegetative cells. Upstream deletions (from -45) reduced the amount of transcription from the promoter. A deletion downstream of the promoter that overlapped a putative downstream minor promoter did not affect transcription from the sigma A promoter, which indicated that the putative downstream promoter is not utilized in vivo. S1 nuclease mapping studies showed that there was a low level of transcription from the subtilisin promoter during the growth phase and that the site of transcription initiation was the same during log and stationary phases. We conclude from these findings that there is only one promoter for the subtilisin gene and that it can be transcribed by the sigma A form of RNA polymerase in vitro.

Bacillus subtilis subtilisin gene (aprE) is expressed from a sigma A (sigma 43) promoter in vitro and in vivo.

INTRODUCTION.............................................................. 568 PROPERTIES OF PROCARYOTIC RPase ............ ......................... 569 Subunit Structure ......................................................... 569 Functions of the Subunits .................................................... 570 Genetics of the RPase Subunits ............................................... 570 Accessory Regulatory Factors ........................ ........................ 572 Recognition Sites on DNA for RPase and Accessory Regulatory Factors ..... ... 573 RPase ROLE IN PROCARYOTIC DEVELOPMENTAL SYSTEMS ...... ......... 575 T4 Phage Development ........................................................ 575 T7 Phage Development ....................................................... 581 Lambda Phage Development ................................................ 582 B. subtilis Phage SP01 Development ............... ........................... 583 B. subtiis Sporulation....................................................... 584 CONCLUSIONS ............................................................... 586 LITERATURE CITED......................................................... 587

https://mmbr.asm.org/content/mmbr/41/3/568.full.pdf

Role of ribonucleic acid polymerase in gene selection in procaryotes.

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Role of ribonucleic acid polymerase in gene selection in procaryotes.