Naegleria fowleri: nfa1 gene knock-down by double-stranded RNAs.
Nfa1 protein expressed by the nfa1 gene that was cloned recently from pathogenic Naegleria fowleri was found in pseudopodia, especially food-cups, and concerned with a mechanism of pathogenicity of N. fowleri. In the present study, N. fowleri nfa1 gene was knocked down using double-stranded RNAs, and the expression of Nfa1 protein was observed. Using synthetic double-stranded RNA of the nfa1 gene in vitro, the nfa1 gene and Nfa1 protein were knocked down about 50.4+/-3.1% and 52+/-2%, respectively. These results suggest that RNA interference (RNAi) may be an effective technique for gene knock-down in N. fowleri trophozoites.
Jung
Suk-Yul
SY
Department of Microbiology, Ajou University School of Medicine, Suwon 443-721, Republic of Korea.
Kim
Jong-Hyun
JH
Lee
Yang-Jin
YJ
Song
Kyoung-Ju
KJ
Kim
Kyongmin
K
Park
Sun
S
Im
Kyung-il
KI
Shin
Ho-Joon
HJ
D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics* / Q000502:physiology; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D004587:Electrophoresis, Agar Gel; D005786:Gene Expression Regulation; D016848:Naegleria fowleri / Q000235:genetics*; D015800:Protozoan Proteins / Q000235:genetics* / Q000502:physiology; D034622:RNA Interference / Q000502:physiology*; D012330:RNA, Double-Stranded / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D000953:Antigens, Protozoan; C460560:Nfa1 protein, Naegleria fowleri; D015800:Protozoan Proteins; D012330:RNA, Double-Stranded