关键词: 致倦库蚊, 细胞系, DNA扩增指纹图谱, 内转录间隔区, 分子鉴定

Abstract: 【Aim】 This study aims to establish and molecularly identify a cell line based on tissues from the newly hatched larvae of the mosquito Culex pipiens quinquefasciatus (Diptera: Culicidae). 【Methods】 The shredded tissues of the newly hatched larvae of the parathion-resistant strain Shengui of C. pipens quinquefasciatus were cultured in M199 complete medium supplemented with 20% heatinactivated fetal bovine serum, and the primary cells were obtained. The primary cells were treated with trypsin and subcultured into a cell line. The tissue origin of the cell line was identified by observing its biological characteristics under a microscope and comparing DNA amplification fingerprinting and sequencing the ribosome DNA internal transcribed spacer. 【Results】 The cell line Cxq-1 was well maintained in M199 complete medium and subcultured continually for 12 months. After being frozen in liquid nitrogen, Cxq-1 was successfully resuscitated for several times. DNA amplification fingerprinting polymerase chain reaction clearly distinguished Cxq-1 from two other insect cell lines. Sequencing of ribosome DNA internal transcribed spacer of Cxq-1 indicated that its rDNA-ITS1 and rDNA-ITS2 sequences has higher than 99% identity with those of C. pipens quinquefasciatus registered in the GenBank, confirming the origin of Cxq-1. 【Conclusion】 The newly established cell line Cxq-1 from newly hatched larvae of C. pipens quinquefasciatus has been confirmed to be a cell line of this mosquito. This cell line provides an important platform for studying the molecular mechanisms of insecticide resistance, mosquito-borne viruses and parasites in the future.

Key words: Culex pipiens quinquefasciatus, cell line, DNA amplification fingerprinting, internal transcribed spacer, molecular identification