Tags

Type your tag names separated by a space and hit enter

In vitro packaging of bacteriophage phi 29 DNA restriction fragments and the role of the terminal protein gp3.
J Mol Biol 1989; 209(1):91-100JM

Abstract

Restriction fragments of bacteriophage phi 29 DNA-gp3 (DNA-gene product 3 complex) were packaged in a completely defined in vitro system that included purified proheads, the DNA packaging protein gp16 and ATP. Both left and right end DNA-gp3 fragments were packaged in this system, in contrast to the oriented and selective packaging of left end DNA-gp3 fragments in extracts; left ends could be packaged quantitatively in the defined system, while the packaging efficiency of right ends was generally about threefold lower. In addition, certain internal (non-end) DNA fragments were packaged at efficiencies of about 10% to 15%. Digestion of the gp3 with trypsin or proteinase K reduced the packaging of whole-length DNA by a factor of 2 or 4, respectively, and removal of the gp3 from whole-length DNA or end fragments with piperidine reduced packaging to the level of internal fragments. Though the terminal protein gp3 was non-essential for DNA translocation in the defined system, it stimulated packaging of left and right end fragments, and stabilized packaging of the left end. The packaging of end and internal DNA fragments of the related phage M2Y into phi 29 proheads was similar to that of phi 29 DNA fragments, and certain fragments of lambda DNA were packaged at the efficiency of the internal phi 29 DNA fragments. Selective packaging of DNA-gp3 left ends was restored by the addition of bacterial cell extracts or glycerol to the defined system, and these packaging conditions discriminated between phi 29 and M2Y DNAs that have distinct terminal proteins.

Authors+Show Affiliations

Department of Microbiology, University of Minnesota, Minneapolis 55455.No affiliation info available

Pub Type(s)

Journal Article
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

2530357

Citation

Grimes, S, and D Anderson. "In Vitro Packaging of Bacteriophage Phi 29 DNA Restriction Fragments and the Role of the Terminal Protein Gp3." Journal of Molecular Biology, vol. 209, no. 1, 1989, pp. 91-100.
Grimes S, Anderson D. In vitro packaging of bacteriophage phi 29 DNA restriction fragments and the role of the terminal protein gp3. J Mol Biol. 1989;209(1):91-100.
Grimes, S., & Anderson, D. (1989). In vitro packaging of bacteriophage phi 29 DNA restriction fragments and the role of the terminal protein gp3. Journal of Molecular Biology, 209(1), pp. 91-100.
Grimes S, Anderson D. In Vitro Packaging of Bacteriophage Phi 29 DNA Restriction Fragments and the Role of the Terminal Protein Gp3. J Mol Biol. 1989 Sep 5;209(1):91-100. PubMed PMID: 2530357.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - In vitro packaging of bacteriophage phi 29 DNA restriction fragments and the role of the terminal protein gp3. AU - Grimes,S, AU - Anderson,D, PY - 1989/9/5/pubmed PY - 1989/9/5/medline PY - 1989/9/5/entrez SP - 91 EP - 100 JF - Journal of molecular biology JO - J. Mol. Biol. VL - 209 IS - 1 N2 - Restriction fragments of bacteriophage phi 29 DNA-gp3 (DNA-gene product 3 complex) were packaged in a completely defined in vitro system that included purified proheads, the DNA packaging protein gp16 and ATP. Both left and right end DNA-gp3 fragments were packaged in this system, in contrast to the oriented and selective packaging of left end DNA-gp3 fragments in extracts; left ends could be packaged quantitatively in the defined system, while the packaging efficiency of right ends was generally about threefold lower. In addition, certain internal (non-end) DNA fragments were packaged at efficiencies of about 10% to 15%. Digestion of the gp3 with trypsin or proteinase K reduced the packaging of whole-length DNA by a factor of 2 or 4, respectively, and removal of the gp3 from whole-length DNA or end fragments with piperidine reduced packaging to the level of internal fragments. Though the terminal protein gp3 was non-essential for DNA translocation in the defined system, it stimulated packaging of left and right end fragments, and stabilized packaging of the left end. The packaging of end and internal DNA fragments of the related phage M2Y into phi 29 proheads was similar to that of phi 29 DNA fragments, and certain fragments of lambda DNA were packaged at the efficiency of the internal phi 29 DNA fragments. Selective packaging of DNA-gp3 left ends was restored by the addition of bacterial cell extracts or glycerol to the defined system, and these packaging conditions discriminated between phi 29 and M2Y DNAs that have distinct terminal proteins. SN - 0022-2836 UR - https://www.unboundmedicine.com/medline/citation/2530357/In_vitro_packaging_of_bacteriophage_phi_29_DNA_restriction_fragments_and_the_role_of_the_terminal_protein_gp3_ L2 - https://linkinghub.elsevier.com/retrieve/pii/0022-2836(89)90172-1 DB - PRIME DP - Unbound Medicine ER -