Reference gene validation for quantification of gene expression during final oocyte maturation induced by diethylstilbestrol and di-(2-ethylhexyl)-phthalate in common carp

Final oocyte maturation is the key step to successful spawning and fertilization.Quantitative real-time PCR(q PCR) is the technique of election to quantify the abundance of functional genes in such study. Reference gene is essential for correct interpretation of q PCR data. However, an ideal universal reference gene that is stable under all experimental circumstances has not been described. Researchers should validate their reference genes while performing q PCR analysis. The expression of 6 candidate reference genes: 18 s r RNA,28 s r RNA, Cathepsin Z, Elongation factor 1-α, Glyceraldehyde-3-phosphate dehydrogenase andβ-actin were investigated during final oocyte maturation induced by different compounds(DES and DEHP) in common carp(Cyprinus carpio). Four softwares(Bestkeeper, ge Norm,Norm Finder and Ref Finder) were used to screen the most stable gene in order to evaluate their expression stability. The results revealed that EF1α was highly stable expressed when final oocyte maturation was induced by DES, while gapdh was the most stable gene when final oocyte maturation was induced by DEHP. Stable expressed reference gene selection is critical for all q PCR analysis to get accurate target gene m RNA expression information.     

己烯雌酚与四溴双酚A对小球藻生长效应的影响（Effects of Diethylstilbestrol and Tetrabromobisphenol A on Growth of Chlorella pyrenoidosa）

采用化学品对藻类毒性测试的标准试验方法,分别研究了己烯雌酚和四溴双酚A单组分对小球藻生长及光合色素含量的影响,并用相加指数法评价了己烯雌酚和四溴双酚A双组分对小球藻的联合毒性效应.结果表明,藻细胞密度和光合色素含量随己烯雌酚和四溴双酚A浓度增加而明显下降,且均表现出较好的浓度-效应关系,其对小球藻的96h-EC50值分别为0.0248mg·L-1和0.0455mg·L-1;两者对小球藻96h的联合毒性效应为拮抗作用.     

气相色谱-串联质谱法测定水中己烯雌酚残留

采用乙酸乙酯提取水样中己烯雌酚残留,经BSTFA/TMCS衍生化后,用GC-MS/MS测定,通过优化试验条件,使该方法在0.05 mg/L~1.00 mg/L范围内线性良好,方法检出限为146 ng/L。实际水样2个质量浓度水平的加标回收率为90.8%和91.9%,5次测定结果的RSD分别为2.2%和2.1%。