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PMID: 6304623 Published · ppublish English Journal Article

Targeted random mutagenesis: the use of ambiguously synthesized oligonucleotides to mutagenize sequences immediately 5' of an ATG initiation codon.

Nucleic acids research ·Vol. 11 ·No. 10 ·1983-05-25 ·Pages 3113-21

Matteucci MD, Heyneker HL

Abstract

The nine base pairs immediately 5' of the initiation codon for the bovine growth hormone (BGH) structural gene have been mutagenized. The mutagenesis method employs the ligation of an ambiguously synthesized oligonucleotide duplex into a previously engineered gap in an expression plasmid for BGH. The mutation method, coupled with hybridization screening, is efficient at isolating 1 and 2 base pair changes within the targeted region. The E. coli cultures harboring the mutant plasmids were assayed for relative levels of BGH expression. The most notable result is the varied effect of substitution of G into the mRNA at various positions in this region.

MeSH Terms
Animals Base Composition Base Sequence Cattle Codon/genetics DNA Restriction Enzymes Escherichia coli/genetics Genes/drug effects Growth Hormone/genetics Mutation Nucleic Acid Hybridization Oligodeoxyribonucleotides/pharmacology Oligonucleotides/pharmacology Plasmids RNA, Messenger/genetics
Chemicals
Codon Oligodeoxyribonucleotides Oligonucleotides RNA, Messenger Growth Hormone DNA Restriction Enzymes
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Matteucci M D
Heyneker H L
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19 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/11.10.3113
Published
1983-05-25
Pages
3113-21
Language
English
Region
England
NLM ID
0411011
PMCID
PMC325952
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/11.10.3113 PMC Full Text View journal details
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