摘要
【目的】探索环境内分泌干扰物壬基酚(Nonylphenol,NP)对贵州草海鲫鱼(Carassius auratus)Spo11基因表达的影响。【方法】通过PCR方法克隆实验鱼Spo11基因,利用生物信息学方法分析SPO11蛋白的氨基酸序列、理化性质和结构特点;并设置1个对照组和低、中、高共3个不同剂量NP处理组,用NP质量浓度分别为0,25,50和100μg·L^(-1)的水体浸浴实验鱼28d后,检测各组实验鱼Spo11基因的表达情况。【结果】Spo11基因cDNA序列长度为1 471bp(GenBank登录号:MF66470),包括66bp的5′非翻译区、编码383个氨基酸的1 152bp的开放阅读框和253bp的3′非翻译区。预测SPO11蛋白质相对分子质量为43.23kDa,等电点IP为6.89,主要由亮氨酸、丝氨酸和缬氨酸组成。该蛋白是含有信号肽的不稳性亲水蛋白,二级结构以α-螺旋和无规则卷曲为主。与对照组实验鱼的Spo11基因相对表达量相比,中剂量和高剂量的NP处理组实验鱼Spo11基因相对表达量均有所升高,与前者的差异均具有统计学意义(p<0.05)。【结论】环境内分泌干扰物NP可以影响贵州草海鲫鱼Spo11基因的表达。
[Purposes]To explore the effect of environmental hormone Nonylphenol（NP）on the Spo11 gene expression of the crucian carp（Carassius auratus）in Caohai lake of Guizhou Province.[Methods]The cDNA of Spo11 gene was cloned by PCR in crucian carp.And its physico-chemical properties and structure characteristics were analyzed by bioinformatics.Spo11 gene expression were detected after the fish were treated with NP mass concentrations of 0,25,50,and 100μg·L^（-1） for 28 days.[Findings]It was shown that the sequence of Spo11 gene was 1 471 bp（GenBank ID：MF66470）in length,containing 66 bp 5′UTR,1 152 bp ORF,and 253 bp 3′UTR.The predicted molecular weight was 43.23 kDa,and the value of PI was 6.89.Proteins are mainly made up of leucine,serine and valine.And the protein is an unstable hydrophilic protein containing signal peptide.Its secondary structures were shown to be mainlyα-helix and random coil.Compared with the control group,the relative expression of Spo11 gene were significantly higher in medium and high concentration groups after NP treatment（p〈0.05）.[Conclusions]It is shown that the environmental endocrine disruptor nonylphenol can affect the Spo11 gene expression of crucian carp in Caohai lake of Guizhou Province.
作者
董然然
陈修云
王岗屹
吕昌乾
DONG Ranran;CHEN Xiuyun;WANG Gangyi;LU Changqian(College of Animal Science,Guizhou University,Guiyang 550025,Chin)
出处
《重庆师范大学学报（自然科学版）》
CAS
CSCD
北大核心
2018年第4期25-30,F0002,共7页
Journal of Chongqing Normal University:Natural Science
基金
贵州省科技厅联合基金(黔科合LH字[2015]7678)
教育部重点实验室开放课题(No.FFRD-2015-01)
关键词
贵州草海鲫鱼
Spo11基因
基因克隆
生物信息学
壬基酚
crucian carp in Caohai lake of Guizhou Province
Spo11 gene
gene cloning
bioinformatics
nonylphenol
