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中华绒螯蟹蜕皮抑制激素基因全长cDNA克隆和重组表达 被引量:7
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作者 孙妍 张亦陈 +5 位作者 刘逸尘 王雪惠 王宇凡 耿绪云 孙金生 杨卫军 《水生生物学报》 CAS CSCD 北大核心 2011年第2期210-217,共8页
根据实验室分离自中华绒螯蟹(Eriocheir sinensis)的一种蜕皮抑制激素(Molting-inhibiting hormone,MIH)N端氨基酸测序结果设计简并引物,采用RACE方法,首次从中华绒螯蟹眼柄中克隆到蜕皮抑制激素基因全长cDNA(Es-MIH,GenBank登录号:DQ34... 根据实验室分离自中华绒螯蟹(Eriocheir sinensis)的一种蜕皮抑制激素(Molting-inhibiting hormone,MIH)N端氨基酸测序结果设计简并引物,采用RACE方法,首次从中华绒螯蟹眼柄中克隆到蜕皮抑制激素基因全长cDNA(Es-MIH,GenBank登录号:DQ341280),该基因全长为1457 bp,开放阅读框为330 bp,编码110个氨基酸(含有35个氨基酸的信号肽);其成熟肽包含C7-C44、C24-C40和C27-C53三个二硫键,有典型的CHH家族结构域。该cDNA编码的氨基酸序列与地蟹(Gecarcinus lateralis)MIH同源性最高,达到了85%。Northern杂交和半定量RT-PCR显示蜕皮间期成体蟹仅在眼柄中有MIH基因表达,提示该基因的表达具有一定组织特异性。利用pCR T7/NT TOPO TA系统重组表达MIH成熟肽,纯化的重组蛋白得率为0.3 g/L,纯化产物经质谱鉴定为中华绒螯蟹MIH。研究解决了CHH家族神经肽在机体中的表达量少,直接纯化较难的问题,为深入研究MIH的作用机制和在生产上控制中华绒螯蟹蜕皮和生长奠定了基础。 展开更多
关键词 中华绒螯蟹 蜕皮抑制激素基因 组织表达 重组表达 纯化
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重叠延伸拼接法从基因组DNA中获得Ers-MIH1基因的编码区
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作者 郭豫杰 鲁维飞 +1 位作者 韩立强 杨国宇 《河北渔业》 2006年第10期13-15,48,共4页
蜕皮抑制激素(molt-inhibiting hormone,MIH)产生于端髓X-器官(MTXO),储存在甲壳动物眼柄的窦腺中,抑制Y-器官蜕皮素的分泌,参与调控甲壳动物的生长和发育等重要的生理活动。提取中华绒螯蟹肌肉组织中的基因组DNA,以此为模板,采用重叠... 蜕皮抑制激素(molt-inhibiting hormone,MIH)产生于端髓X-器官(MTXO),储存在甲壳动物眼柄的窦腺中,抑制Y-器官蜕皮素的分泌,参与调控甲壳动物的生长和发育等重要的生理活动。提取中华绒螯蟹肌肉组织中的基因组DNA,以此为模板,采用重叠延伸拼接法(gene splicing by overlap extension,SOE)将中华绒螯蟹蜕皮抑制激素1Eriocheir sinensismolt-inhibiting hormone-1(Ers-MIH1)成熟肽基因的编码区连接起来,克隆到pMD18-T载体中,随机挑取3个阳性克隆测序,测序结果与已知的Ers-MIH1基因编码区的序列相一致。从基因组DNA中克隆该基因的方法有效地解决了取材不便给研究中华绒螯蟹蜕皮抑制激素基因所带来的限制等问题。 展开更多
关键词 中华绒螯蟹 基因组DNA 蜕皮抑制激素基因1 重叠延伸拼接法
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Effects of Periodical Salinity Fluctuation on the Growth, Molting, Energy Homeostasis and Molting-Related Gene Expression of Litopenaeus vannamei 被引量:1
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作者 ZHANG Dan GUO Xiantao +1 位作者 WANG Fang DONG Shuanglin 《Journal of Ocean University of China》 SCIE CAS 2016年第5期911-917,共7页
To determine the response of Litopenaeus vannamei to periodical salinity fluctuation, a 30-day experiment was conducted in laboratory. In this experiment, two salinity fluctuation amplitudes of 4(group S4) and 10(grou... To determine the response of Litopenaeus vannamei to periodical salinity fluctuation, a 30-day experiment was conducted in laboratory. In this experiment, two salinity fluctuation amplitudes of 4(group S4) and 10(group S10) were designed. The constant salinity of 30(group S0) was used as the control. Levels of shrimp growth, molting frequency(MF), cellular energy status(ATP, ADP and AMP), as well as the expression of genes encoding molt-inhibiting hormone(MIH), crustacean hyperglycemic hormone(CHH), ecdysteroid-regulated protein(ERP), and energy-related AMP-activated protein kinase(AMPK) were determined. The results showed that periodical salinity fluctuation significantly influenced all indicators except MF which ranged from 13.3% in group S10 to15.4% in group S4. In comparison with shrimps cultured at the constant salinity of 30, those in group S4 showed a significant elevation in growth rate, food conversion efficiency, cellular energy status, ERP and MIH gene transcript abundance, and a significant reduction in CHH and AMPK transcript abundance(P < 0.05). However, salinity fluctuation of 10 only resulted in a significant variation in MIH and CHH gene expression when compared to the control(P < 0.05). According to our findings, L. vannamei may be highly capable of tolerating salinity fluctuation. When ambient salinity fluctuated at approx. 4, the increased MF and energy stores in organisms may aid to promoting shrimp growth. 展开更多
关键词 salinity shrimp fluctuation transcript periodical AMPK ranged Molting abundance Periodical
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