条纹密鲈卵黄蛋白原基因的克隆与检测  

Cloning,sequence analysis and detection of vitellogenin cDNA from Colisa fasciata

作　　者：梁岳[1] 方展强[1] 

机构地区：[1]华南师范大学生命科学学院广东省高等学校生态与环境科学重点实验室,广东广州510631

出　　处：《水产学报》2011年第9期1281-1292,共12页Journal of Fisheries of China

基　　金：国家科技支撑计划项目(2009BADB2B0401-02);广东省科技计划项目(2009B030600006)

摘　　要：采用RT-PCR方法克隆并分析了条纹密鲈卵黄蛋白原(VTG)和β-肌动蛋白(β-actin)cDNA部分序列。获得的VTG cDNA序列片段长3 464 bp,全部处于编码区,编码1 150个氨基酸;β-actin cDNA序列片段长1 253 bp,编码375个氨基酸。使用活体与离体的实验方法,检测了VTG mRNA转录情况,并以此评价雌二醇(E2)、辛基酚(OP)、镉(Cd2+)和全氟辛烷磺酸类化合物(PFOS)引起的雌激素效应。结果显示,E2和OP在活体和离体实验中均能剂量依赖性地诱导VTG mRNA表达。Cd2+仅在活体实验低剂量组诱导VTG mRNA表达,PFOS在活体和离体实验的各个浓度组均未见显著的VTG mRNA表达。结果表明,所诱导的雌激素效应强弱的排列顺序为E2>OP>Cd2+。Cd2+的雌激素效应活体和离体实验的结果不一致,推测Cd2+引起雌激素效应的机理可能和E2不同。条纹密鲈VTG cDNA对环境激素敏感,适合作为监测环境激素的生物标志物。Using RT-PCR,the partial length vitellogenin（VTG）cDNA sequence and β-actin cDNA sequence of Colisa fasciata were cloned.VTG cDNA sequence contains 3 464 bp nucleotides and encodes 1 150 amino acids.β-actin cDNA sequence contains 1 253 bp nucleotides and encodes 375 amino acids.In vivo and in vitro methods were employed to investigate VTG mRNA expression under exposure to estradiol（E2）,octylphenol（OP）,cadmium（Cd2＋）and perfluorooctane sulfonates（PFOS）and evaluate the estrogenic activity.The results showed that E2 and OP could induce VTG mRNA expression in dose-dependent way by in vivo and in vitro test.Cd2＋ could induce VTG mRNA expression only in the low dose by in vivo test,but VTG mRNA expression was not observed in PFOS groups by in vivo and in vitro test.The results indicated that the strength of estrogenic effects was in the order E2OPCd2＋.Cd2＋ estrogenic effects in vivo and in vitro results are inconsistent,suggesting that the mechanism of Cd2＋ induced effects of estrogen and E 2 may be different.The results also indicated that VTG cDNA of C.fasciata is very sensitive to environmental hormone and very suitable to be a biomarker for monitoring the environmental hormones.

关 键 词：条纹密鲈 卵黄蛋白原基因 克隆与表达 环境激素 

分 类 号：S917[农业科学—水产科学]