siRNA of schistosoma japonicum katsurada SjLwr gene and application thereof
A technology for schistosomiasis and schistosomiasis, which is applied in the fields of molecular biology and biomedicine, and can solve problems such as drug resistance, affecting clinical application effects, and large first-pass effects
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Embodiment 1
[0027] Example 1: In vivo screening of small RNAs
[0028] 1. Method steps
[0029] 1.1 siRNA molecules and wxya Preparation of primers for quantitative real-time PCR of genes
[0030] Find schistosomes on the NCBI website (http: / / www.ncbi.nlm.nih.gov / ) Lwr Gene sequence (FN321279.1). according to Lwr Expression of gene open reading frame in Schistosoma japonicum, design and synthesis of three pairs of dsRNA to produce Lwr Gene-specific inhibitory effects, affecting Lwr gene expression. Simultaneously, a pair of unrelated control dsRNA (NCsiRNA) was synthesized.
[0031] S1 dsRNA: sense: 5'-GCGUUAUUCAAUGCCGCAUTT-3'(SEQ NO.1)
[0032] anti-sence: 5'-AUGCGGCAUUGAAUAACGCTT-3'(SEQ NO.2)
[0033] S2 dsRNA: sense:5'-GCUACAGUCCAACCACAAATT-3'(SEQ NO.3)
[0034] anti-sence: 5'-UUUGUGGUUGGACUGUAGGTT-3'(SEQ NO.4)
[0035]S3 dsRNA: sense:5'-GCUACUUGACUCCCUACUUTT-3'(SEQ NO.5)
[0036] anti-sence:5'-AAGUAGGGAGUCAAGUAGCTT-3'(SEQ NO.6)
[0037] NC dsRNA:sense:5'-UUCUCCGAACG...
Embodiment 2
[0050] Embodiment 2: RNA interference experiment
[0051] 1. Method steps
[0052] 1.1 dsRNA injection
[0053] The experiment consisted of control group, unrelated control group (NC group) and S2 dsRNA treatment group, with three mice in each group. 11 days, 15 days, 19 days, 23 days, 27 days, 31 days, 35 days and 39 days after infection, 200ul DEPC water, 200ul NC dsRNA (8µg) and 200ul SjLwr gene S2 dsRNA (8ug ), a total of 8 times.
[0054] 1.2 Hatch rate of liver worm eggs
[0055] After dissecting the mouse, take the liver and weigh it, add dechlorinated water to make it up to 20 ml, mix it with a homogenizer, take 4 ml of the crushed liquid and add it to the chicken heart bottle (add dechlorinated water to the neck of the chicken heart bottle 3-5 cm above), put cotton on the liquid surface of the chicken heart bottle, add 8 ml of dechlorinated water to the cotton, incubate at 26-27 ℃ for 2 h, transfer 8 ml of the upper layer of cotton to a 15 ml conical centrifuge A...
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