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目的克隆我国重要蚊媒的防御素基因并对其序列进行分析。方法分别提取埃及伊蚊、白纹伊蚊、致乏库蚊、中华按蚊的基因组DNA和总RNA。根据国外已发表的防御素基因序列设计、合成引物,进行PCR和RT-PCR。结果分别从埃及伊蚊、白纹伊蚊、中华按蚊基因组DNA中扩增出预期片段,将片段切胶纯化后进行序列分析并与已发表的防御素基因比较,显示埃及伊蚊和白纹伊蚊的扩增片段与已发表的防御素基因序列有很高的同源性,而中华按蚊的扩增片段则与已发表的防御素基因序列的同源性较低,且不具备特征性的6 个半胱氨酸序列。结论克隆出埃及伊蚊和白纹伊蚊的防御素基因,蚊虫防御素基因的同源性高低与其遗传分类距离有一定的平行关系。
Objective To clone the defensin genes of important mosquito vectors in China and analyze their sequences. Methods The genomic DNA and total RNA of Aedes aegypti, Aedes albopictus, Culex pipiens pallens, Anopheles sinensis were extracted respectively. According to foreign published defensin gene sequence design, synthesis of primers, PCR and RT-PCR. Results The expected fragments were amplified from Aedes aegypti, Aedes albopictus and Anopheles sinensis respectively. The fragments were gel purified and sequenced. Compared with the published defensin genes, the results showed that Aedes aegypti and Aedes aegypti The amplified fragment of Aedes mosquitoes had high homology with published defensin gene sequences, whereas the amplified fragment of Anopheles sinensis had lower homology with published defensin gene sequences and had no characteristic Sexual six cysteine sequences. Conclusion The defensin genes of Ae. Aegypti and Aedes albopictus were cloned, and the homology of mosquito defensin genes was in parallel with their genetic distance.