Loading...
REPAIR-RESISTANT NUCLEOSIDE ANALOGUES IN OLIGONUCLEOTIDE-DIRECTED MUTAGENESIS
Jiricny, Josef Ubasawa, Aiko Wood, Steven G.
Jiricny, Josef
Ubasawa, Aiko
Wood, Steven G.
Citations
Altmetric:
Advisors
Editors
Other Contributors
Issue Date
1987
Submitted date
2023-04-19
Files
Loading...
PDF
Adobe PDF, 5.11 MB
Other Titles
Abstract
2'-Deoxytubercidin (a) and 2'-deoxy-7-deazainosine (b) were used as analogues of deoxyadenosine and deoxyinosine respectively for the introduction of transition mutations into the a-peptide fragment of the lacZ gene encoded in the filamentous bacteriophage Ml13mp9. Using conventional methodology of oligonucleotide-directed mutagenesis, the number of desired point mutants was very low (2-6%) for both T+C and C+T transitions, due to the efficient removal of purine/pyrimidine mismatches by the repair machinery of E. coli. Following the incorporation (via synthetic oligodeoxyribonucleotides) of the purine nucleoside analogues a and b into the mismatch sites, the yield of desired mutants was increased approximately tenfold.
Citation
Chemical synthesis in molecular biology, 43 ff
Publisher
DOI
PubMed ID
PubMed Central ID
Additional Links
Embedded video
Type
Book chapter
conference paper
conference paper
Language
en
Description
Series/Report no.
GBF Monographs, Vol. 8
ISSN
0930-4320
EISSN
ISBN
ISMN
Gov't Doc #
Sponsors
License
Attribution-NonCommercial-ShareAlike 4.0 International