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- Newsgroups: bionet.software
- Path: sparky!uunet!zaphod.mps.ohio-state.edu!darwin.sura.net!welchgate.welch.jhu.edu!danj
- From: danj@welchgate.welch.jhu.edu (Dan Jacobson)
- Subject: Re: software for silent mutagenesis
- Message-ID: <1992Dec30.215116.24937@welchgate.welch.jhu.edu>
- Organization: Johns Hopkins Univ. Welch Medical Library
- References: <1992Dec30.185104.9506@alw.nih.gov>
- Date: Wed, 30 Dec 1992 21:51:16 GMT
- Lines: 170
-
- In article <1992Dec30.185104.9506@alw.nih.gov> usdin@helix.nih.gov (Ted Usdin) writes:
- >Weiner and Scheraga (CABIOS, vol 5 pp191-198, 1989) described a Macintosh
- >program (Gene.Design.?) to assist in introducing restriction sites into DNA without changing the amino
- >acid sequence. Does anyone know where I can get a copy of this, or a similar
- >program that runs on a Macintosh?
- >
- >Thanks for any help.
- >
-
- There is a similar program which runs on a PC called OligoMutantMaker.
- It is available by anonymous ftp from:
-
- Host fly.bio.indiana.edu
-
- Location: /molbio/ibmpc
- FILE -rw-r--r-- 8540 May 14 1991 oligo.doc
- FILE -rw-r--r-- 231882 May 14 1991 oligo.uue
-
-
- I am including some information about the program below.
-
-
- Happy New Year,
-
-
- Dan Jacobson
-
- danj@welchgate.welch.jhu.edu
-
-
- ==============================================================================
-
-
- OligoMutantMaker (version 1.0)
- Copyright 1987 Kevin Beadles, David Canter, Arnold Berk
- ("readme.doc" added 8/26/88, 2/24/89 by Spencer Yeh)
-
-
- INTRODUCTION
-
- OligoMutantMaker simplifies the designing and screening of
- oligonucleotide-directed single amino acid substitution experiments by
- searching for nucleotide sequences which introduce a restriction
- endonuclease recognition sequence into the codon substitution site of the
- mutant.
- The program utilizes the redundancy of the genetic code to generate all
- possible nucleotide sequences for a given amino acid substitution
- (including nucleotide sequences in which silent mutations are introduced
- into the 5' and/or 3' codons immediately adjacent to the substitution site)
- and determines whether any restriction endonuclease recognition
- sequences are present. Any nucleotide sequence containing a restriction
- site is displayed/printed along with relevant information about the site
- such as its restriction enzyme(s), the random frequency of the enzyme's
- recognition sequence(s), the prototype of the enzyme, isoschizomers of the
- enzyme, and the unit cost of the enzyme from various biochemical
- suppliers.
-
-
- CONTACT ADDRESS
-
- For questions about the program or suggestions for future improvements
- please contact:
-
- Kevin Beadles
- 1044 1/2 Shrader St.
- San Francisco, CA 94117
-
- tel.: (415) 759-0148
- (Kevin will call you back collect if he must return your call.)
-
-
-
- SYSTEMS SUPPORTED
-
- An IBM compatible computer running MS-DOS is needed.
-
-
-
-
-
-
- PROGRAM FILES before de-ARCing
-
- README.DOC This documentation file.
- OLIGO.UUE Archived and uuencoded file for both monochrome
- and color executable versions, and associated data
- files. (234 Kb).
-
-
-
- PROGRAM FILES after de-ARCing (Approx. 350 Kb total)
-
- README.DOC This documentation file
- BWMUTANT.COM Executable file for monochrome monitors
- CMUTANT.COM Executable file for color monitors
- CUTTERS.DAT Binary database of enzyme cleavage and availability data
- CODONID.DAT Standard genetic code
- [ENZYME.TXT] This file is created everytime the program is run.
- It contains a copy of the results of the last analysis.
-
-
- DOCUMENTATION
-
-
- The program is internally documented with help messages.
-
-
- (....)
-
- ADDITIONAL INFORMATION ABOUT THE PROGRAM
-
- Restriction Site Location:
- OligoMutantMaker searches for restriction sequences which have at
- least one nucleotide in codon #0 (i.e. the codon to be substituted). Since
- the longest recognition sequence (GGCCNNNNNGGCC--Sfl I) spans thirteen
- nucleotides, it is necessary to enter 27 nucleotides (twelve nucleotides on
- either side of the three nucleotides which comprise codon #0) into the
- program in order for it to function.
-
- "Silent" Mutations:
- The program utilizes the redundancy of the genetic code to generate
- all possible nucleotide sequences for the given amino acid substitution in
- codon #0 (i.e. the codon to be substituted). In addition to searching for
- restriction sites which match these nucleotides sequences, the program
- also examines those nucleotide sequences in which "silent" mutations have
- been introduced into codon #-1 and codon #+1 (i.e. the codons immediately
- upstream--or 5'--and downstream--or 3'--from codon #0, respectively)
- and determines whether any additional restriction sites are created. Only
- those "silent" mutations which can be created by a single substitution in
- the nucleotide closest to codon #0 (i.e. the third--or "wobble"--nucleotide
- of codon #-1 and the first nucleotide of codon #+1) are examined.
- Therefore, only five nucleotides (centered upon the middle nucleotide of
- codon #0) are subject to substitution.
-
-
- Abbreviations used in OligoMutantMaker:
- standard three letter amino acid abbreviations
- STP: stop codon (UAA, UAG, or UGA)
- standard one letter nucleotide abbreviations
- N: any nucleotide
- m: methylated nucleotide
- /: cleavage site
- standard restriction endonuclease abbreviations
- BMB: Boehringer Mannheim Biochemicals
- BRL: Bethesda Research Laboratories
- NEB: New England Biolabs
- USBC: United States Biochemical Corperation
- u: unit(s)
- c/u: cents/unit
-
-
- Miscellaneous Notes:
- OligoMutantMaker recognizes 164 different restriction sites which are
- are cleaved by 126 different restriction enzymes.
- Price comparisons are based upon the smallest low concentration order
- available from each supplier.
- OligoMutantMaker does not search for cleavage specificities generated
- by methylating DNA with a selected restriction endonuclease (see NEB
- 1986/87 Catalog p32 & p123 for further details).
- OligoMutantMaker does not note enzyme specificities resulting from
- star activity (NEB 1986/87 Catalog p86 & p121).
-
-
-
- Bibliography:
- Boehringer Mannheim Biochemicals, 1987/88 (Catalog)
- Bethesda Research Laboratories, Catalog & Reference Guide (1985)
- New England Biolabs, Catalog 1986/87
- United States Biochemical Corporation, Enzymes & Reagents for
- Molecular Biology 1987
-