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为了解日本囊对虾(Marsupenaeusjaponicus)性腺发育及分化的分子机制,本研究采用改良的引物退火控制技术克隆到了共8个精巢、卵巢的差异表达基因.其中4个为已知基因,分别为凝血酶敏感蛋白基因、皮质棒蛋白基因、增殖细胞核抗原基因和泛素缀合酶基因;4个为未知基因,分别命名为MjACP3-T、MjACP4一T、MjACPl5一T和MjACP4一O基因.采用实时定量PCR方法分析了4个未知基因在性腺发育过程的表达谱,结果表明:MjACP3一T、MjACP4.T和MjACPl5,T基因在日本囊对虾精巢和卵巢中的表达量有显著差异,且在各个时期精巢的表达量均高于卵巢,说明这3个基因可能在精巢的发育中起到某些重要作用.卵巢MjACP4一O基因的表达量变化较大,第5期卵巢的相对表达量达到最高(为1.38),而其他各个阶段的变化较大,在第4期卵巢的相对表达量最低(为0.03).在精巢中,MjACP4一O基因的表达量随着精巢逐渐发育成熟呈下降趋势.定量PCR结果与引物退火控制技术的结果有着很好的一致性.研究结果表明,引物退火控制技术是一种简单高效的克隆差异表达基因的技术.采用该技术所克隆到的8个精巢、卵巢的差异表达基因,为进一步了解日本囊对虾性腺发育分化的分子机制提供了基础材料. 相似文献
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甲壳动物高血糖激素家族在调控甲壳动物的变态发育、生殖、蜕皮、血糖平衡及体内的渗透压平衡等方面起着重要的作用,甲壳动物高糖激素(CHH)是甲壳动物高血糖激素家族中的重要成员.通过Tail-PCR方法克隆获得拟穴青蟹(Scylla paramamosain)chh基因的近端启动子区序列,从翻译起始位点(ATG)起,共657 bp.生物信息学分析结果显示,转录起始位点位于翻译起始位点(ATG)上游第74个碱基(A),在转录起始位点(A)上游31 bp处存在TATA box;核心启动子区位于-41~+9 bp之间,包含在线软件预测的转录起始位点;潜在的转录因子结合位点包括Oct-1、C/EBPalp、GATA-1、Sp1、Ap-1、NF-κappa B、NF-1、RAP1、HNF-3等;目前获得的chh基因启动子区中没有Cp G岛.成功构建了chh基因4个启动子不同长度片段C1—C4,分别位于-583~+205、-388~+205、-286~+205、-46~+205 bp.含启动子不同长度片段的表达载体瞬时转染HEK293T细胞和Sf9细胞后的活性分析表明,在-286~-46、-583~-388 bp区域可能存在负调控基因表达的转录因子结合位点,在-388~-286 bp区域可能存在正调控基因表达的转录因子结合位点. 相似文献
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We investigated the effect of tributyltin (TBT) exposure on the concentration of malondialdehyde (MDA) and the activity levels
of the superoxide dismutase (SOD), catalase (CAT), and acid and alkaline phosphatase (ACP and AKP) enzymes in the small abalone,
Haliotis diversicolor supertexta. We collected samples of the hepatopancreas and hemolymph 2, 6, 24, 48, 96, and 192 h after exposure to 0.35 μg (Sn)/L TBT.
In the hepatopancreas, ACP activity was significantly higher in animals exposed to TBT 2, 24, and 96 h post-exposure compared
with the control animals. AKP activity was also higher after 2 h, but SOD and CAT activity was unchanged. The concentration
of MDA in the hemolymph was significantly higher than the control animals 2 and 6 h post-exposure. In the hemolymph of animals
exposed to TBT, ACP activity was significantly lower than in the control animals 192 h post-exposure, whereas AKP activity
was significantly lower 2 and 192 h post-exposure. Hemolymph SOD activity and levels of MDA were significantly lower than
in the control animals 24 h after exposure but significantly higher after 96 h. Our results demonstrate that exposure to TBT
cause rapid changes in ACP and AKP activity as well as altering the concentration of MDA in the hepatopancreas and hemolymph.
SOD and CAT do not appear to be involved in the detoxification of TBT in the hepatopancreas of small abalone. 相似文献
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