知识与财富的链接
中国明对虾卵黄蛋白原mRNA基因在卵巢和肝胰腺中都有表达。采用实时荧光定量PCR方法测定在卵巢不同发育时期卵巢和肝胰腺这两种组织的卵黄蛋白原mRNA的表达水平,提取组织总RNA,设计特异性引物进行荧光定量PCR扩增,对卵巢发育的各个阶段,卵原细胞增殖期、卵黄发生前期、初级卵黄发生期、次级卵黄发生期和消退期的卵黄蛋白原mRNA表达进行相对定量分析。结果表明:在卵巢中,卵黄蛋白原mRNA在前3个阶段随着卵巢的发育,表达量不断增加,分别为0.04、0.23和1.03,次级卵黄发生期开始减少到0.16,消退期卵黄蛋白原mRNA表达量又增加到1.00。而在肝胰腺,前4个阶段随着卵巢发育,表达量逐步增加,分别为0.01、0.07、0.19、和1.29,至消退期,表达量减少到0.74。研究亮点:使用实时荧光定量的方法,分析了中国明对虾卵黄蛋白原mRNA基因在卵巢发育的不同时期,卵巢和肝胰腺组织中的相对表达水平。选择敏感性和特异性较好的实验方法,在前人定性研究的基础上开始相对定量的研究,清晰地阐述了卵巢和肝胰腺中卵黄蛋白原mRNA基因的表达差异。
The vitellogenin mRNA gene was expressed in both ovary and hepatopancreas of shrimp Fenneropenaeus chinensis.In this paper,the expression of vitellogenin mRNA in the ovary and hepatopancreas at different ovarian development stages,including multiplication oogoniium,previtellogenesis,primary vitellogenesis,secondary vitellogenesis and declining stages was determined by real-time fluorescence quantitative PCR.The total RNA was extracted from the ovary and hepatopancreas and the target gene was amplified by real-time fluorescence quantitative PCR with special primers.The results show that in ovary,the expression of mRNA increased from multiplication oogoniium to primary vitellogenesis stage.The relative expression levels were 0.04,0.23 and 1.03 respectively,but decreased at secondary vitellogenesis stage to 0.16.However,in declining stage it was 1.00.In hepatopancreas,the expression of mRNA increased from multiplication oogoniium to secondary vitellogenesis stage.The relative expression levels were 0.01,0.07,0.19 and 1.29 respectively,and it decreased to 0.74 at decling stage.
