Specific protein gene and protein of female blood fluke and their application
A technology for specific protein and schistosome female, applied in the field of biological genes, can solve the problem of no public report on the specific protein of female adult Schistosoma japonicum, etc., and achieve the effect of preventing the spread
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Embodiment 1
[0031] Bioinformatic analysis of proteins specific to female adult worms of Schistosoma japonicum.
[0032] 1. Screening of specific proteins of Schistosoma japonicum female adults: using expressed sequence tag (EST) data to analyze specifically expressed proteins. The genes specifically expressed by females relative to males: 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 of females must be 3 times or more than that of males. After the screening of these three criteria, the specific expression genes of female adults were obtained.
[0033] 2. Homology and similarity analysis: BLASTP comparison of the specific protein sequence of Schistosoma japonicum female adult to the non-redundant database (Non-redundant, NR) of GenBank (http: / / www.ncbi.nlm.nih.gov / ) Right; Schistosoma japon...
Embodiment 2
[0035] RT-PCR experiments verified the expression of specific proteins in Schistosoma japonicum female adults.
[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 the homogenizer and homogenize for several minutes; transfer the homogenized liquid into an RNase-free centrifuge tube, add...
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