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目的对淡色库蚊不同龄期幼虫中肠氨肽酶N(Aminopeptidase N,APN)的表达水平进行分析。方法应用反转录-聚合酶链反应(RT-PCR)克隆淡色库蚊APN基因的全长c DNA序列,利用实时荧光定量PCR(q RT-PCR)分析该基因在不同龄期幼虫中肠的表达情况。结果克隆获得淡色库蚊APN基因,该基因全长1 383 bp,编码460个氨基酸,与Gen Bank收录的致倦库蚊(XM001862270.1)序列的同源性为100%。q RT-PCR表明Cp APN基因在各个龄期均有表达,其中在4龄期的表达量最高,3龄期最低,1、2和4龄期的表达量差异无统计学意义(F=2.465,P=0.250),与3龄期幼虫的差异有统计学意义(F=49.191,P=0.000),分别为其表达量的3.5、3.7和4.0倍。结论Cp APN基因在淡色库蚊不同龄期幼虫中的表达水平存在差异,为进一步探讨淡色库蚊APN的功能奠定了基础。
Objective To analyze the expression of aminopeptidase N (APN) in larvae of Culex pipiens pallens at different instars. Methods The full-length c DNA sequence of APN gene of Culex pipiens pallens was cloned by reverse transcription-polymerase chain reaction (RT-PCR). Real-time quantitative PCR (q RT-PCR) Express the situation. Results The APN gene of Culex pipiens pallens was cloned. The gene is 1 383 bp in length and encodes a protein of 460 amino acids. It has 100% homology with the GenBank accession number of Culex pipiens quinquefasciatus (XM001862270.1) in Gen Bank. q RT-PCR showed that Cp APN gene was expressed at all ages, with the highest expression in the fourth instar, lowest in the third instar, and no significant difference in the first and the fourth instar (F = 2.465 , P = 0.250). The difference was statistically significant (P = 0.000, P = 0.000) with the 3rd instar larvae, which were 3.5, 3.7 and 4.0 fold respectively. Conclusion The expression level of Cp APN gene in different instar larvae of Culex pipiens pallens is different, which lays the foundation for further study on the function of APN in Culex.