Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases · 2005
[RNA interference to the expression of peroxiredoxin-related genes in Trichomonas vaginalis].
Аннотация на английскомСтатьяНазвание и аннотация: Перевод в очереди
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Zhang Jia-xin, Fu Yu-cai, Xu Xiao-yuan, Wu Tong-jian, Cao Feng-ling
Аннотация
OBJECTIVE: To inhibit the expression of the target genes of peroxiredoxin (Prx) and thioredoxin reductase (TrxR) by RNA interference and evaluate its effect on the growth of Trichomonas vaginalis.
METHODS: Genomic DNA was extracted from cultured Trichomonas vaginalis with phenol-chloroform method and was transcribed to double stranded RNA (dsRNA). Short interference RNAs (siRNA, 21-23 bp) synthesized by digestion of dsRNA with RNase III and purified through filter cartridge, were transfected into the cells in three groups (A, B and C) to degrade the target genes of Prx, TrxR and Prx+TrxR through siPORT lipid, respectively, and the untransfected was selected as a control (group D). The levels of Prx and TrxR mRNA were determined 24 h and 48 h post-transfection by relative quantitative RT-PCR, and the growth of Trichomonas vaginalis was estimated under microscope 36 h post-transfection.
RESULTS: Trichomonas vaginalis mRNA levels of Prx and TrxR decreased. Though the cell activity showed no significant difference (P > 0.05) in four groups as expected, a difference existed (P < 0.01) between the groups in the average of vells (7.2 x 10(7)/L, 14.2 x 10(7)/L, 3.8 x l0(7) L and 20.3 x 10(7)/L in groups A, B, C and D respectively).
CONCLUSIONS: RNA interference inhibits the expression of the genes of Prx and TrxR and extended Trichomonas vaginalis cells cycle considerably, but showed no influence on the cell activity.
METHODS: Genomic DNA was extracted from cultured Trichomonas vaginalis with phenol-chloroform method and was transcribed to double stranded RNA (dsRNA). Short interference RNAs (siRNA, 21-23 bp) synthesized by digestion of dsRNA with RNase III and purified through filter cartridge, were transfected into the cells in three groups (A, B and C) to degrade the target genes of Prx, TrxR and Prx+TrxR through siPORT lipid, respectively, and the untransfected was selected as a control (group D). The levels of Prx and TrxR mRNA were determined 24 h and 48 h post-transfection by relative quantitative RT-PCR, and the growth of Trichomonas vaginalis was estimated under microscope 36 h post-transfection.
RESULTS: Trichomonas vaginalis mRNA levels of Prx and TrxR decreased. Though the cell activity showed no significant difference (P > 0.05) in four groups as expected, a difference existed (P < 0.01) between the groups in the average of vells (7.2 x 10(7)/L, 14.2 x 10(7)/L, 3.8 x l0(7) L and 20.3 x 10(7)/L in groups A, B, C and D respectively).
CONCLUSIONS: RNA interference inhibits the expression of the genes of Prx and TrxR and extended Trichomonas vaginalis cells cycle considerably, but showed no influence on the cell activity.