蚂蟥wpHSP90基因的克隆及不同温度下的表达分析  

Cloning and expression analysis of wpHSP90 gene from Whitmania pigra at different temperatures

作　　者：张会宁 郭巧生[1] 史红专[1] 陈盼盼 郭丽媛 禤汉美 ZHANG Hui-ning;GUO Qiao-sheng;SHI Hong-zhuan;CHEN Pan-pan;GUO Li-yuan;XUAN Han-mei(Institute of Chinese Materia Medica,Nanjing Agricultural University,Nanjing 210095,China;School of Pharmacy,Wannan Medical College,Wuhu 241001,China)

机构地区：[1]南京农业大学中药材研究所,江苏南京210095 [2]皖南医学院药学院,安徽芜湖241001

出　　处：《中国中药杂志》2023年第22期6051-6057,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81673538)。

摘　　要：HSP90是广泛存在的分子伴侣,参与多种细胞行为,在生殖细胞的减数分裂过程中发挥重要作用,但尚不清楚在雌雄同体蚂蟥Whitmania pigra性腺发育中的作用。为探究HSP90对蚂蟥生殖细胞发育的影响,该研究对蚂蟥wpHSP90基因进行了克隆,生物信息学分析以及表达量测定。结果表明,克隆得到的wpHSP90全长2 592 bp,其中开放阅读框(ORF)长为2 373 bp,编码790个氨基酸。预测wpHSP90蛋白在丝氨酸、苏氨酸和酪氨酸残基上有85个磷酸化修饰位点。结构域预测和多序列比较结果显示,wpHSP90含有HSP90的2个保守结构域,与水蛭Helobdella robusta的同源性最高,序列相似度为80.72%。RT-qPCR结果表明,在12~28℃,5月龄蚂蟥性腺中wpHSP90的相对表达量与温度呈正相关,其中雌性腺中的表达量明显高于雄性腺,且与精卵巢生殖细胞发育趋势一致。综上,wpHSP90可能参与调控蚂蟥精卵巢生殖细胞的发育,尤其卵细胞的发育。该研究可为蚂蟥性腺发育机制的进一步研究提供参考。HSP90 is a widely distributed molecular chaperone that participates in a variety of cellular processes and plays an important role in the meiosis of germ cells.However,its role in the gonadal development of hermaphroditic Whitmania pigra is not yet clear.To explore the effect of HSP90 on the germ cell development of Wh.Pigra,this study cloned the wpHSP90 gene,performed bioinformatics analysis,and measured its expression levels.The results showed that the cloned wpHSP90 was 2 592 bp in length,with an open reading frame(ORF) of 2 373 bp,encoding 790 amino acids.Prediction analysis revealed 85 phosphorylation modification sites on serine,threonine,and tyrosine residues of the wpHSP90 protein.Structural domain prediction and multiple sequence alignment results showed that wpHSP90 contained two conserved domains of HSP90 and exhibited the highest homology with Helobdella robusta,with a sequence similarity of 80.72%.RT-qPCR results showed that the relative expression level of wpHSP90 in the gonads of 5-month-old Wh.pigra was positively correlated with temperature within the range of 12 ℃ to 28 ℃.The expression level in the female gonads was significantly higher than in the male gonads and correlated with the trend of germ cell development in the ovaries and testes.In conclusion,wpHSP90 may be involved in regulating the development of germ cells,particularly oocytes,in Wh.pigra.This study provides a reference for further research on the gonadal development mechanism in Wh.pigra.

关 键 词：蚂蟥 HSP90 克隆 表达分析 性腺发育 

分 类 号：Q953[生物学—动物学]