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目的克隆鉴定白纹伊蚊气味结合蛋白基因OBP68,了解该基因在白纹伊蚊不同发育时期表达量的差异。方法提取幼虫、蛹、雄蚊、未吸血雌蚊总RNA,用RT-PCR扩增OBP68基因特异片段,克隆于pMD 18-T载体并测序鉴定;用半定量RT-PCR分析白纹伊蚊不同发育时期OBP68基因mRNA表达量差异。结果从白纹伊蚊扩增出OBP68基因,测序分析克隆白纹伊蚊OBP68基因序列与GenBank注册号FJ04086.1的白纹伊蚊OBP68基因同源性99%,与埃及伊蚊的OBP68基因的同源性为71%。白纹伊蚊OBP68基因在白纹伊蚊发育不同时期皆有表达,以幼虫阶段表达较弱,蛹、雄蚊、雌蚊表达量增高,且后3个时期表达量差异无统计学意义(P>0.05)。结论从白纹伊蚊成功扩增出OBP68基因,该基因在白纹伊蚊不同发育时期均有表达。
Objective Clone identification of Aedes albopictus odorant binding protein gene OBP68, understand the gene expression in different developmental stages of Aedes albopictus differences. Methods The total RNA of larvae, pupae, male mosquitoes and non-blood-sucking female mosquitoes was extracted. The specific fragment of OBP68 gene was amplified by RT-PCR and cloned into pMD 18-T vector and sequenced. Semiquantitative RT-PCR was used to analyze Aedes albopictus Differences of OBP68 gene mRNA expression during development. Results The OBP68 gene was amplified from Aedes albopictus. The sequence analysis of the OBP68 gene of Ae. Albopictus was 99% homologous with the OBP68 gene of Aedes albopictus in GenBank accession number FJ04086.1, Homology was 71%. The expression of OBP68 gene of Aedes albopictus was expressed at different stages of Aedes albopictus development. The larval expression of OBP68 gene was low, and the expression levels of pupae, male mosquito and female mosquito increased, and there was no significant difference in the expression of OBP68 (P > 0.05). Conclusion OBP68 gene was successfully amplified from Aedes albopictus and expressed in different developmental stages of Aedes albopictus.