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The repressor gene (c) of the Streptomyces temperate phage ϕc31: Nucleotide sequence, analysis and functional cloning (1988)

Sinclair, Robert B. ; Bibb, Mervyn J.

Springer

Molecular genetics and genomics 213 (1988), S. 269-277

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ISSN: 1617-4623

Keywords: Recombinant DNA ; Helix-turn-helix motifs ; In vitro transcription-translation ; Phage immunity ; Exonuclease III deletions

Source: Springer Online Journal Archives 1860-2000

Topics: Biology

Notes: Summary The nucleotide sequence of the 3.4 kb SphI-G fragment that contained the repressor gene (c) of the temperate Streptomyces phage ϕc31 was determined. Analysis of this sequence revealed a large open reading frame with protein coding character and sequence changes in c gene point and deletion mutants identified this as the coding region of the repressor. Two of the mutants studied had undergone deletions of 1.1 kb and 1.4 kb that had occurred across short direct repeats of 6 bp and 11 bp, respectively. Coupled in vitro transcription-translation experiments using the cloned SphI-G fragment and Streptomyces lividans cell free extracts identified a protein product of approximately 72 kDa, in close agreement with that predicted from the nucleotide sequence. A strongly predicted helix-turn-helix motif that may be involved in DNA binding occurred towards the carboxy-terminus of the amino acid sequence. Initial attempts to clone the SphI-G fragment in Streptomyces failed; using information gained from the sequence analysis a smaller segment of this DNA fragment was cloned in S. lividans and conferred immunity to a clear plaque mutant (c1) of ϕc31.

Type of Medium: Electronic Resource

URL: http://dx.doi.org/10.1007/BF00339591

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The preparation of cross-section specimens for transmission electron microscopy (1984)

Bravman, John C. ; Sinclair, Robert

New York, NY : Wiley-Blackwell

Journal of Electron Microscopy Technique 1 (1984), S. 53-61

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ISSN: 0741-0581

Keywords: Cross-section specimen ; Thin films ; Interfaces ; Life and Medical Sciences ; Cell & Developmental Biology

Source: Wiley InterScience Backfile Collection 1832-2000

Topics: Natural Sciences in General

Notes: The structure and chemistry of thin solid films are best studied by transmission electron microscopy (TEM) when they are viewed in cross-section - that is, when the surface normal of the film is made perpendicular to the electron beam. In this orientation, the substrate, the thin film layers, and the interfaces between them can be imaged either simultaneously or individually. Further, information from each of these regions remains distinct from that obtained from the others, eliminating the problems of superimposition that are a consequence of viewing a layered structure in the conventional manner (i.e., parallel to the surface normal). A technique for fabricating TEM specimens that can be viewed in cross-section is described here. Although the majority of our work is with silicon-based materials, the technique can be readily adapted to the study of other systems.

Additional Material: 8 Ill.