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Mutational analysis of the specific priming signal essential for DNA replication of the broad host‐range plasmid RSF1010
Author(s) -
Honda Yoichi,
Akioka Tae,
Takebe So,
Tanaka Katsunori,
Miao Dengming,
Higashi Akihisa,
Nakamura Toshiaki,
Taguchi Yoshitomo,
Sakai Hiroshi,
Komano Tohru,
Bagdasarian Michael
Publication year - 1993
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(93)81534-7
Subject(s) - primase , plasmid , primer (cosmetics) , priming (agriculture) , biology , mutant , dna , base pair , nucleotide , dna replication , genetics , microbiology and biotechnology , chemistry , gene , polymerase chain reaction , reverse transcriptase , germination , botany , organic chemistry
To analyze the RSF1010‐specific priming mechanism, a library of randomly mutagenized ssiA sequences was constructed by chemical synthesis using mixed nucleotide phosphoramidites. Synthetic ssiA sequences with the single base‐substitutions were assayed for the SSI activity in E. coli JM109 expressing RepB' primase. It was demonstrated that the activity of ssiA was damaged markedly by single base‐substitutions within the possible stem‐loop structure and its 3'‐flanking region. It is conceivable that these domains are critical in recognition and primer synthesis by RepB' primase.