A modified kanamycin-resistance cassette to facilitate two-codon insertion mutagenesis.
A kanamycin-resistance (KmR) cassette was modified to allow its introduction as a blunt-ended DNA fragment into target plasmids. Subsequent excision of the KmR cassette with ApaI and religation of the plasmid leaves behind six base pairs (5'-GGGCCC) that result in the insertion of two amino acids in the target protein.
Chen
W J
WJ
Department of Microbiology and Immunology, SUNY-Health Science Center, Brooklyn 11203-2098.
Gross
L
L
Joho
K E
KE
McAllister
W T
WT
D001483:Base Sequence; D003062:Codon; D004247:DNA; D012321:DNA-Directed RNA Polymerases / Q000235:genetics; D007613:Kanamycin Resistance / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional / Q000379:methods*; D010957:Plasmids; D015183:Restriction Mapping; D013604:T-Phages / Q000235:genetics; D014764:Viral Proteins
D003062:Codon; D014764:Viral Proteins; D004247:DNA; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases