【摘要】 目的鉴定和克隆嗜人按蚊雌蚊差异表达基因并对基因序列进行分析。方法以嗜人按蚊为试验模型,根据差异表达基因EST序列(GenBankTM接受号:EX916923)设计引物,采用实时定量PCR(real-time PCR)和RACE(rapid amplifica-tion of cDNAend)技术对雌蚊差异表达基因进行鉴定并扩增其cDNA序列。结果利用F=2-△△CT公式计算出雌蚊差异表达基因在雌蚊和雄蚊中的表达比值为267.49;差异表达基因全长mRNA序列为364bp,开放阅读框(ORF)编码的氨基酸序列经BLAST分析与致倦库纹(Culexquinque fasciatus)结构蛋白(tectin)及其他物种蛋白序列具有相似性。序列发送至NCBI获得接受号:FJ907236。结论该基因为雌蚊差异表达的基因,其全长mRNA序列的获得为进一步研究其生物学功能奠定了基础。更多还原

【Abstract】 To identify the gene differentially expressed in female Anopheles anthropophagus and to analyze its gene sequence,this gene amplified by PCR was identified by real-time PCR and its cDNA was then amplified with rapid amplification of cDNA ends(RACE) technology.It was found that the expression ratio of the female differentially expressed gene in female and male mosquitoes was 267.49 according to the formula F=2-⊿⊿CT.The size of mRNA of the gene was 364 bp,and the amino acid sequence deduced from the open reading frame(ORF) was found to be similar to the sequence of tectin protein of Culex quinquefasciatus and proteins of other species.The mRNA sequence of this gene was submitted to NCBI with a accession number of FJ907236.This gene may provide a foundation for further studies on the biological functions of mosquitoes.更多还原