CAJ | 학술논문

目的：分离、鉴定和分析埃及伊蚊性别决定基因Transformer 2（Aaetra2）。方法通过生物信息学方法与分子生物学方法获得了Aaetra2基因的全长。通过与已知物种tra2的比较，分析Aaetra2的基因结构特点与分子进化特征，通过RT-PCR方法验证Aaetra2的时间与组织表达特点。结果分离并鉴定Aaetra2-α与Aaetra2-β两个基因，它们位于染色体不同部位，通过选择性启动子的剪切策略进行自我调控，转录多种mRNA。埃及伊蚊TRA2蛋白具有保守的RRM、RS1、RS2和linker功能结构域区。Aaetra2表达方式无时间特异性的，从卵持续性表达至成虫阶段，且无性别特异性表达与组织特异性表达。结论 Aaetra2具有保守的昆虫性别决定基因tra2的功能结构域与表达特征，对其深入研究将为其最终应结合释放携带显性致死基因的昆虫技术应用于蚊虫的防制提供理论依据。
목적：분리、감정화분석애급이문성별결정기인Transformer 2（Aaetra2）。방법통과생물신식학방법여분자생물학방법획득료Aaetra2기인적전장。통과여이지물충tra2적비교，분석Aaetra2적기인결구특점여분자진화특정，통과RT-PCR방법험증Aaetra2적시간여조직표체특점。결과분리병감정Aaetra2-α여Aaetra2-β량개기인，타문위우염색체불동부위，통과선택성계동자적전절책략진행자아조공，전록다충mRNA。애급이문TRA2단백구유보수적RRM、RS1、RS2화linker공능결구역구。Aaetra2표체방식무시간특이성적，종란지속성표체지성충계단，차무성별특이성표체여조직특이성표체。결론 Aaetra2구유보수적곤충성별결정기인tra2적공능결구역여표체특정，대기심입연구장위기최종응결합석방휴대현성치사기인적곤충기술응용우문충적방제제공이론의거。
Objective To isolate, identify and analyze the sex-determining gene Transformer 2 (Aaetra2) of the major vector mosquito Aedes aegypti. Methods tBLASTn program, RT-PCR and RACE methods were used to obtain the full-length cDNA of Aaetra2. Multiple alignments of nucleotide and amino acid sequences were conducted, and the different domains in tra2 protein were indentified. RT-PCR of the total RNA extracted from different tissue from the mosquitoes in different developmental stages was performed using specific primers. Results Two genes, namely Aaetra2-α and Aaetra2-β, were identified in different supercontig locations. The multi-transcripts were expressed by means of alternative promoters or terminators. The different domains in tra2 protein were defined as RS-rich N-terminal region, RNA recognition motif-RRM, linker region, and RS-rich C-terminal region. Both Aaetra2-α and Aaetra2-β showed sustained expression throughout the developmental stages of Ae.aegypti, and in all the tissues without a sex specificity. Conclusion Aaetra2 gene has multiple isoforms and is mapped to multiple locations in the genome. Aaetra2 has conservative functional domains of the sex-determining gene tra2. For Ae.agypti, Aaetra2 shows the potential as a new target for release of insects carrying a dominant lethal (RIDL) technology based on transgenic mosquitoes.