海洋科学2016，Vol.40Issue(9)：23-31,9.DOI:10.11759//hykx20150201004
诸氏鲻虾虎鱼卵黄蛋白原基因全长cDNA的克隆及表达
Full-length cDNA cloning and expression analysis of vitellogenin gene inMugilogobius chulae
摘要
Abstract
To explore the tissue distribution of the vitellogenin (Vg) gene inMugilogobius chulae and the effect of the pollutant 17β-estradiol on maleM. chulae, the full length of vitellogenin (Vg) cDNA in M. chulae was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE) tech-niques using total RNA extracted from the liver of female fish. Meanwhile, the expression of Vg mRNA ofM. chu-lae was determined by real-time fluorescent quantitative PCR. The full length ofM. chulae’s Vg cDNA sequence contains 5 067 bp nucleotides, with a complete open reading frame (ORF) of 4 992 bp, which encodes 1 664 amino acid proteins with a deduced protein molecular weight of 186.2 ku. The ORF contains signal peptides and a polyserine domain. The real-time RT-PCR result showed thatM. chulae’s Vg mRNA was primarily detected in the liver. Furthermore, Vg mRNA expression levels at different time points of E2 treatment of male fish were deter-mined.M. chulae’s Vg mRNA expression levels reached the peak on the third day and then the expression levels declined on the seventh day. These results indicated that the full length of Vg cDNA in M. chulae was cloned suc-cessfully. This study suggests that Vg in maleM. chulae can be considered as a valid biomarker for offshore envi-ronmental monitoring of estrogenic substances.关键词
诸氏鲻虾虎鱼(Mugilogobius chulae)/卵黄蛋白原/cDNA全长/荧光定量PCR/17-雌二醇Key words
Mugilogobius chulae/vitellogenin/full-length cDNA/quantitative real-time RT-PCR/17β-estradiol分类
资源环境引用本文复制引用
余露军,蔡磊,李舸,陈小曲,陈琳,李建军..诸氏鲻虾虎鱼卵黄蛋白原基因全长cDNA的克隆及表达[J].海洋科学,2016,40(9):23-31,9.基金项目
国家科技支撑计划项目(2013BAK11B902) （2013BAK11B902）
广东省科技计划项目(2011B010500021,2012B040302006)[Foundation：National Key Technology Support Program (2013BAK11B902) （2011B010500021,2012B040302006）
Science and Technology Program of Guangdong Province (2011B010500021,2012B040302006) （2011B010500021,2012B040302006）
