ABSTRACT
Objective To explore the expression characteristics of cytochrome P450(CYP)candidate genes(CYP6M3, CYP6Y1,CYP6P5,CYP4H14,CYP4G17,CYP12F16)in Anopheles sinensis.Methods The samples were collected from dif-ferent developmental stages(eggs,larvae,pupae and adult mosquitoes(females and males)),and different tissues(salivary glands,malpighian tubes,midguts,ovaries,and fat bodies)of An.sinensis and the female adult mosquitoes exposed by differ-ent insecticide doses(0,1.25,3.75,6.25,12.5μg/bottle)and time points(0,5,15,30,60 minutes),then the total RNA was extracted. Quantitative reverse transcription PCR(qPCR)assay was used to analyze the relative expressions of six CYP genes in An.sinensis at different developmental stages,tissues and different insecticide exposure doses and time.Results The expressions of CYP6M3 and CYP6Y1 in the male adult mosquitoes were the highest,the expression of CYP6M3 gene in the males was 35.1 times higher than that in the females,the expression of CYP6Y1 in the males was 61.4 times higher than that in the females;the expression level of CYP4H14 in the larvae was the lowest,the expression of CYP4H14 in the females was 22.5 times higher than that in the fourth instar larvae.The expressions of candidate CYP genes in different tissues of An.sinensis were significantly different,the expression of CYP6M3 in the malpighian tubule was 38.9 times higher than that in the ovary,the ex-pression of CYP6Y1 in the fatbody was 9.1 times higher than that in the ovary,the expression of CYP4G17 was 4.6 times higher than that in the ovary,and the expression of CYP12F16 was 4.4 times higher thanthat in the ovary.The exposure to different in-secticide doses and time showed some induction effects in the expressions of candidate CYP genes,which affected the expres-sions of candidate CYP genes in An.sinensis.Conclusion The expression of candidate CYP genes varies significantly in differ-ent developmental stages and various tissues of An.sinensis,and exposure to deltamethrin at various doses and time points af-fects CYP genes expression in An.sinensis.
ABSTRACT
Objective To establish the MALDI-TOF-MS technique platform for detecting Cytochrome P450 gene polymor-phismfrom of patients.Methods Collected 53 EDTA anticoagulation peripheral blood samples from October 2013 to June 2014 in Peking Union Medical College Hospital.The whole genomic DNA was extracted from patients’peripheral blood. Used MALDI-TOF-MS to identify the SNP polymorphism of CYP2C9*2(rs1799853),CYP2C9*3(rs1057910),CYP2C19*2(rs4244285),CYP2C19*3(rs4986893),CYP2C19*4(rs28399504),CYP2C19*5(rs56337013)and CYP2C19*17 (rs12248560).To verify the above SNP polymorphism by Sanger sequencing method.Results The MALDI-TOF-MS could perform 53 samples on two cytochrome gene and 7 SNP locus simultaneously.In all the 53 patients,25 AG,6 AA and 22 GG genotypes were identified in CYP2C19*2(rs4244285),the allele frequency of A genotype was 34.9%.4 AG and 49 GG genotypes were identified in CYP2C19*3 (rs4986893),the allele frequency of A genotype was 3.8%.5 CA and 48 AA gen-otypes were identified in CYP2C9*3 (rs1057910),the allele frequency of C genotype was 4.7%.No mutation loci were i-dentified in CYP2C9*2 (rs1799853),CYP2C19*4 (rs28399504),CYP2C19*5 (rs56337013)and CYP2C19*17 (rs12248560).All the identification data were confirmed by Sanger sequencing.The coincidence rate was 100%.Conclusion The MALDI-TOF-MS technique platform for the cytochrome enzyme SNP was established.This platform has high throughput and accurate characteristics.It has important clinical application value for the treatment of personalized medicate.