ASTM E1493-92(1998)
(Guide)Standard Guide for Identification of Bacteriophage M13 or Its DNA
Standard Guide for Identification of Bacteriophage M13 or Its DNA
SCOPE
1.1 This guide covers the identification of bacteriophage M13 used in biotechnology.
1.2 There are many variants of M13 that have been developed specifically for cloning technology. These variants have foreign DNA inserted into the M13 genome, causing the M13 to differ in size and genotype.
1.3 If the M13 is to be used to construct a recombinant molecule, then the criteria described in Section 6 should be used to characterize the newly made DNA.
General Information
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Standards Content (Sample)
NOTICE: This standard has either been superseded and replaced by a new version or
withdrawn. Contact ASTM International (www.astm.org) for the latest information.
Designation: E 1493 – 92 (Reapproved 1998)
AMERICAN SOCIETY FOR TESTING AND MATERIALS
100 Barr Harbor Dr., West Conshohocken, PA 19428
Reprinted from the Annual Book of ASTM Standards. Copyright ASTM
Standard Guide for
Identification of Bacteriophage M13 or Its DNA
This standard is issued under the fixed designation E 1493; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (e) indicates an editorial change since the last revision or reapproval.
INTRODUCTION
This guide covers the identification of bacteriophage M13 or its DNA and was developed by Task
Group E48.02.03 on viruses. The objective is to describe laboratory characterization procedures that
would be sufficient to verify that a biological preparation believed to contain M13 (or M13 DNA) for
use in any step of a biotechnology process actually does contain this bacteriophage (or its DNA).
This guide assumes a basic knowledge of microbiology and molecular biology.
1. Scope 4. Background Information about M13 Bacteriophage
1.1 This guide covers the identification of bacteriophage 4.1 M13 is a filamentous bacteriophage that infects male
+
M13 used in biotechnology. (F F or Hfr) Escherichia coli. The phage particles contain
1.2 There are many variants of M13 that have been devel- circular single-stranded DNA, 6407 nucleotides in length,
oped specifically for cloning technology. These variants have coated with the protein product of the M13 gene 8.
foreign DNA inserted into the M13 genome, causing the M13 4.2 The phage attaches to a receptor at the end of the F pilus.
to differ in size and genotype. The infecting single-stranded DNA ( + strand) replicates in the
1.3 If the M13 is to be used to construct a recombinant cell: a complementary (−) strand is synthesized, resulting in a
molecule, then the criteria described in Section 6 should be double-stranded, replicative form (RF). Using the RF as a
used to characterize the newly made DNA. template, both strands can be replicated, increasing the copy
number of the RF to about 20 to 40 per cell. Late in infection
2. Terminology
(+) strands are preferentially produced and packaged into
2.1 Definitions:
phage particles for export from the cell.
2.1.1 alpha complementation—the ability of a short amino- 4.3 M13 infection is not lethal to Escherichia coli. Bacte-
terminal fragment (alpha fragment) of b-galactosidase to form
riophage DNA is continually replicated and packaged, causing
a functional complex with the carboxyl terminal fragment a decrease in the growth rate of the host. The “plaque” seen
(omega fragment).
upon infection by M13 is the result of an area of decreased
2.1.2 F—an F factor that contains a portion of the E. coli growth rate, not actually cell killing.
genome.
4.4 M13 has extensive sequence homology to bacterioph-
2.1.3 F factor—an episome of E. coli. Encoded on it are the ages f1 and fd, differing in only a few bases.
functions necessary to produce an F pilus.
4.5 M13 is used in biotechnology primarily as a vector into
2.1.4 F pilus—a protrusion on E. coli that is necessary for which foreign DNA can be cloned. Commonly used M13
mating. The F pilus also contains the receptor for phage M13.
variants are the M13 mp series. These M13 bacteriophages
2.1.5 multiple cloning site—DNA that contains several have a portion of the E. coli lac operon with a multiple cloning
contiguous restriction enzyme recognition sites; also called a
site within a truncated lacZ gene.
polylinker. 4.5.1 A portion of the E. coli lac operon containing the
3-prime end of the lacI gene, the lac promoter and operator,
3. Significance and Use
and the alpha complementation portion of the lacZ gene is
3.1 This guide is intended for use in a biotechnology
laboratory w
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