REPAIR-RESISTANT NUCLEOSIDE ANALOGUES IN OLIGONUCLEOTIDE-DIRECTED MUTAGENESIS
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Issue Date
1987Submitted date
2023-04-19
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Show full item recordAbstract
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 ffAffiliation
Friedrich Miescher-Institut, PO Box 2543, CH-4002 BaselType
Book chapterconference paper
Language
enSeries/Report no.
GBF Monographs, Vol. 8ISSN
0930-4320Collections
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