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. 1972 Jun;69(6):1555–1559. doi: 10.1073/pnas.69.6.1555

Transfecting Deoxyribonucleic Acid of Bacillus Bacteriophage ϕ29 That Is Protease Sensitive

Hideo Hirokawa 1
PMCID: PMC426747  PMID: 4504368

Abstract

The transfecting activity of Bacillus phage ϕ29 DNA, extracted either by sodium lauroyl sarcosine-phenol or by 2 M perchlorate, was destroyed by treatment with proteolytic enzymes, although these enzymes did not effect transfecting DNAs of SPP1, SPO1, and SP50. These facts suggest that a protein is associated with transfective ϕ29 DNA. Stabilization of protease-resistance during transfection appeared earlier than that of DNaseresistance, indicating that the protein associated with ϕ29 DNA is necessary for initiation of the incorporation of DNA molecules into competent cells. The physical nature of ϕ29 DNA before and after the trypsin treatment was investigated by sucrose and CsCl density gradient centrifugations. The trypsin treatment did not alter the sedimentation rate of the unit ϕ29 DNA; however, it did convert the sedimentation rate of the aggregated material in the untreated DNA to that of the unit ϕ29 DNA. The density of the trypsinized DNA was 0.009 g/cm3 greater than that of the untreated DNA. The possible location of the protein on the DNA is discussed.

Keywords: trypsin; CsCl centrifugation; sucrose gradient centrifugation; SPP1, SPO1, and SP50 bacteriophages

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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