Japanese blood fluke specific sex relevant gene and its coded protein and application
A specific protein, schistosomiasis technology, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problem of no public report on the specific protein of the male adult Schistosoma japonicum, and achieve the effect of preventing the spread
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Embodiment 1
[0031] Bioinformatic analysis of proteins specific to male adults of Schistosoma japonicum.
[0032] 1. Screening of specific proteins of Schistosoma japonicum male adults: using expressed sequence tag (EST) data to analyze specifically expressed proteins. Genes specifically expressed in males relative to females: 1) The significance of the difference must satisfy p<0.05, and this significance analysis is calculated using a special software tool on the Internet; 2) The number of ESTs for each gene must be greater than or equal to 10 ; 3) The number of ESTs in males must be 3 times or more than that in females. After the screening of these three criteria, the specific expression genes of male adults were obtained.
[0033] 2. Homology and similarity analysis: BLASTP comparison of the specific protein sequence of Schistosoma japonicum male adult to the non-redundant database (Non-redundant, NR) of GenBank (http: / / www.nebi.nlm.nih.gov / ) Right; BLASTP homology comparison of the ...
Embodiment 2
[0035] RT-PCR experiment was used to verify the expression of the specific protein in adult Schistosoma japonicum male worms.
[0036] 1. Sample Preparation
[0037] Adult females and males of Schistosoma japonicum were prepared for RNA extraction.
[0038] 2. Total RNA extraction kit
[0039] The RNA extraction reagent uses TRIzol reagent (GIBCO / BRL), which is produced based on the one-step extraction method with acidic phenol. The utensils and water used for RNA extraction must be RNase-free to ensure an RNase-free environment in the experiment.
[0040] 3. RNA extraction steps
[0041] Dry-bake the pestle, homogenizer and other utensils at 200°C for 4 hours to remove RNase and cool down; add liquid nitrogen to pre-cool, take the tissue out of the liquid nitrogen quickly, and grind it into powder; use a spatula to put the tissue into the pre- Add TRIzol reagent to a homogenizer and homogenize for several minutes; transfer the homogenized liquid into an RNase-free centrif...
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