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THE ARRANGEMENTS OF NUCLEOTIDE SEQUENCES IN T2 AND T5 BACTERIOPHAGE DNA MOLECULES.

The genetic map of T4 (and T2) bacteriophage is circular but the DNA molecule that is liberated by phenol extraction is a linear duplex of polynucleotide chains. If the genetic map is related to the physical structure of the DNA molecule, the problem arises as to how a linear molecule can give rise to a circular map. An explanation can be made on the basis that the bacteriophage liberate molecules which have nucleotide sequences which are circular permutations of each other. Thus, markers which are most distant on one molecules are closest together on another. To test this hypothesis, the middles of T2 and T5 DNA molecules were mechanically deleted and the absence of certain nucleotide sequences was tested by "renaturation" or "reannealing" experiments using columns containing denatured DNA immobilized in agar beads. The results indicate that when the middles are deleted from the T5 DNA molecule, some special sequences are removed; whereas, when the middles are deleted from the T2 DNA molecule, no special group of sequences is removed. This would indicate that T2 molecules begin at different points in their nucleotide sequence, while T5 molecules all begin at the same point. It is likely that this permutation of sequences of T2(T4) molecules is related to the circularity of their genetic map.

Bacteriophages↗

THE EFFECT OF TEMPERATURE ON THE FORMATION OF T1 AND T2R BACTERIOPHAGE.

A very rapid variation of yield of T1 and T2r bacteriophage in E. coli at slightly higher than normal temperatures has been observed. T1 phage will develop at 41.2 degrees C but not at 41.7 degrees C. By infecting cells grown on lactose and, therefore, induced to contain beta galactosidase, a technique which indicates when cells have become leaky was worked out. This method shows that at elevated temperatures the enzymatic attack on the cell wall continues to go at a faster rate, while completion of the phage goes more slowly. Thermal constants are given for the processes. Cells at higher temperature, grown on P(32) medium develop incomplete particles capable of combining with phage antibody. This suggests that the process affected by the early leakiness of the cells is the completion of the virus protein coat. Supplementing the medium with casamino acids, phosphate, and ATP causes the "rescue" of phage particles by aiding the formation of the coat. This can be achieved several minutes after the cells have become leaky and may form a useful system for the study of phage development in the presence of analogs.

Adenosine Triphosphate↗

CHROMATOGRAPHIC SEPARATION OF ANNEALED AND ENZYMATICALLY SYNTHESIZED RNA-DNA HYBRIDS.

A procedure is described for the chromatographic detection and isolation of DNA-RNA hybrids on columns of methylated albumin coated on kieselguhr (MAK). Its use is illustrated with both annealed and enzymatically synthesized hybrids. The method has the advantage of a wide range in capacity and resolution and permits actual isolation of the hybrid structure. It is uniquely effective in experiments involving hybridization with small DNA fragments.

Chromatography↗