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本文应用随机引物扩增多态性DNA聚合酶链反应技术筛选出引物Pm,用此引物对淡色库蚊细胞系(CPP-512)、中华按蚊卵巢细胞系(Anso-242)、嗜人按蚊细胞系(Ana-104)、白纹伊蚊细胞株(C6/36)4种蚊细胞基因组DNA进行扩增,得出4个相异的多态性DNA扩增产物图谱,各自的DNA扩增条带的分子量(kb)为CPP-512:0.5、0.7、0.8、0.88、1.02、1.22;Anso-242: 0.5、0.88、1.02、1.13、1.22、1.5、2.1、2.4、2.5;Ana-104:0.64、0.98、1.2、1.28、1.4、1.53、1.8、2.3、2.6、3.3;C6/36:0.66、0.8、0.94、1.4、1.42、1.5、1.52、2.9。实验结果表明,随机引物扩增多态性DNA聚合酶链反应技术是一种简便易行、结果重复性好的先进生物学分类技术,引物Pm能满足同时鉴别3属蚊虫的4种蚊细胞的需要。
In this paper, the primer Pm was screened by random primer-amplified polymorphic DNA polymerase chain reaction (PCR-RFLP). The PCR products were used to detect the expression of Pm in Culex pipiens pallenstris cell line (CPP-512), Anopheles sinensis ovary cell line (Ana-104) and Aedes albopictus cell lines (C6 / 36) were used to amplify the genomic DNA of four mosquito cells. Four different polymorphic DNA amplification product maps were obtained, The molecular weight (kb) of the band was CPP-512: 0.5,0.7,0.8,0.88,1.02,1.22; Anso-242: 0.5,0.88,1.02,1.13,1.22,1.5,2.1,2.4,2.5; Ana-104 : 0.64, 0.98, 1.2, 1.28, 1.4, 1.53, 1.8, 2.3, 2.6, 3.3; The experimental results show that the random primer amplification polymorphic DNA polymerase chain reaction (PCR) is a kind of advanced biological taxonomy with good reproducibility and repeatability. Primer Pm can meet the needs of simultaneous identification of four mosquito cells need.