Lépine G, Ellen R P, Progulske-Fox A
Department of Oral Biology, College of Dentistry, University of Florida, Gainesville, Florida 32610, USA.
Infect Immun. 1996 Apr;64(4):1467-72. doi: 10.1128/iai.64.4.1467-1472.1996.
Targeted insertional mutagenesis was used to construct hagA, hagB, and hagC hemagglutinin mutants of Porphyromonas gingivalis. pJRD215-derived plasmids containing tetA(Q)2 and portions of the targeted genes were conjugated into P. gingivalis. Interruption of the three loci was confirmed by Southern hybridization, sequencing, reverse transcription-PCR, and microtiter hemagglutination assays. No significant differences in hydrophobicity or coadherence to Actinomyces viscosus were detected between the mutants and the wild-type strain.
采用靶向插入诱变技术构建牙龈卟啉单胞菌的hagA、hagB和hagC血凝素突变体。将含有tetA(Q)2和部分靶向基因的pJRD215衍生质粒接合到牙龈卟啉单胞菌中。通过Southern杂交、测序、逆转录PCR和微量滴定板血凝试验证实了这三个位点的中断。在突变体和野生型菌株之间未检测到疏水性或与粘性放线菌共粘附性的显著差异。