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外源基因导入鲤受精卵最佳时期的研究 总被引:7,自引:0,他引:7
本研究以鲤鱼受精卵为材料,通过显微注射技术,在受精卵的不同发育时期把鲤鱼MT启动子基因和大麻哈鱼GH基因导入受精卵中,按孵出的鱼苗数分别计算成活率,再通过斑点杂交和Southern Blot杂交检测整合情况,计算整合率,孵化率最高的为单细胞中期(90.83%),整合率最高的为单细胞后期和囊胚期(40%)。综合研究结果外源基因导入鲤鱼受精卵的最佳时期应选在单细胞后期。 相似文献
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从1985年以来,世界各国先后应用不同外源基因导入虹鳟鱼、鲤鱼、鲶鱼等有经济价值的养殖鱼类。但受精卵发育到何时注基因源射外获得高的成活率(孵化率)和整合率一直是研究人员所关心的问题。法国的Chourout等在1986年就做了虹鳟鱼受精卵不同发育时期注射外源基因与孵化率关系实验。美国的Thoms.T.then等人也做了鲤鱼等受精卵在不同发育时期注射外源基因对其成活率、整合率影响的研究。由于外源基因对受体鱼的整合机理比较复杂,不仅这方面的研究不多,而且各实验室之间的研究结果也很不一致,因此至今对某种鱼类的受精印发育到何时是注射外源基因的最佳时期还没有形成一致的意见。本研究以鲤鱼受精卵为材料,通过显微注射技术,在受精卵的不同发育时期把鲤鱼MT启动因子基因,大麻哈鱼CH基因的融合基因导入受精卵中,按孵出的鱼苗数分别计算成活率,再通过斑点杂交和Southern Blot杂交检查整合情况,计算整合率,以探索外源基因导入鲤鱼受精卵的最佳时期。 相似文献
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《Aquaculture (Amsterdam, Netherlands)》1986,51(2):143-150
A plasmid containing human growth hormone cDNA sequence was injected into cytoplasm of fertilized rainbow trout (Salmo gairdneri Rich.) eggs before first cleavage. Survival rates at hatching (stage of analysis) were high (average: 77%; range: 50–100%). Foreign DNA was detected in pooled and individual embryos either by dot blot or by southern electrophoresis procedures using the labelled plasmid as probe; it comigrated with the high molecular weight DNA and was therefore most likely integrated in the trout genome. Transformation yields were higher with linearized than with circular plasmid (75% versus 40%). These data suggest that transgenic trout can be routinely obtained in large proportions. 相似文献
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ABSTRACT: Thermal tolerance was studied in a rainbow trout strain successively selected through high temperature breeding at 20–27°C since 1966 in Miyazaki Prefecture, Japan. The hatching rate and fry mortality at high temperatures were examined in the selected strain along with normal strains cultured at water temperature of 9–17°C. The hatching rate of embryos fertilized at either 10 or 14°C and subsequently subjected to high temperatures in the blastula or neurula stage of the selected strain, was marginally higher than that of the normal strain counterparts. The upper 50% lethal temperatures (LT50 ) for embryos in the early segmentation, blastula and neurula stages of the selected strain were also higher than those of the normal strain counterparts. Death temperatures and LT50 of fries acclimated to 20°C of the selected strain were significantly higher than those of the normal strains. However, no difference in the critical thermal maximum was detected between the different strains. These results suggest that the selected strain of rainbow trout established by selecting successively for many generations at high temperatures acquired a degree of thermal tolerance. 相似文献
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为探索花(Hemibarbus maculates)(♀)×唇(H.labeo)(♂)远缘杂交的可行性,2009—2011年进行花(♀)×唇(♂)的远缘杂交试验。结果表明:采用常规人工干法授精获得花(♀)×唇(♂)杂交受精卵,受精卵在(20±1)℃水温下孵化,受精率、孵化率和出苗率分别达到89.3%、72.6%和67.9%。杂交子一代仔鱼在受精后68h,30min出膜。经过25~30d饲养,夏花鱼种体长达到3~4cm,成活率66%。试验结果证明花(♀)×唇(♂)杂交子一代受精卵能够正常发育,且鱼苗、鱼种及成鱼都能正常生长发育。 相似文献
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为了探究三疣梭子蟹受精卵的离体孵化技术及效果,本研究先后开展了受精卵块最适分离液种类及作用条件的筛选、分离液处理不同发育期受精卵离体孵化的差异、分离液处理对受精卵卵膜的结构影响,及分离液处理受精卵对孵化后幼体活力的影响实验。结果显示,木瓜蛋白酶是一种较为理想的分离液,在浓度为0.09 g/mL,分离时间30 min时,分离率可达到99%以上;经过分离液处理后的各期受精卵均能孵化出幼体,卵内溞状幼体期离体胚胎孵化率最高,为89.0%±3.3%,未经处理的对照组为70.0%±4.8%;卵裂期离体胚胎孵化率最低,为58.0%±3.9%,对照组为31.0%±2.3%,与对照组相比,处理组的孵化率明显提高。透射电镜结果显示,经过分离液处理的受精卵卵膜结构疏松,且厚度降低,符合处理组孵化率增加这一现象,干露、福尔马林溶液胁迫和行为学测试对不同处理组的幼体进行质量评价的结果显示,处理组和对照组幼体活力无显著差异。研究表明,实验所获得的分离液可以有效提高三疣梭子蟹受精卵的分离率和孵化率,且不影响幼体质量,可为三疣梭子蟹及其他甲壳动物受精卵的离体孵化提供参考。本研究可为三疣梭子蟹的苗种繁育提供新的技术手段,也将为基因编辑辅助育种等技术的实施奠定基础。
相似文献16.
The nonnative rainbow trout Oncorhynchus mykiss has been an unsuccessful invader in North European streams, although it has been widely introduced. Here we studied whether early life history stages (egg incubation and hatching, first overwintering) act as filters for the establishment of hatchery rainbow trout. Survival of hatched alevins was approximately 80%, whereas only 47% of the embryos survived. However, the latter value was impacted by the high number of unfertilized eggs. Correlation coefficients with embryo survival rate and environmental variables (pH and temperature) were statistically insignificant. In the overwintering experiments, the survival of rainbow trout was 93%. The growth was generally slowed during the winter, but in the spring the growth of rainbow trout exceeded that of the native brown trout. Our data demonstrated that the survival and growth of rainbow trout during early life-history stages were relatively high and comparable to those of the native brown trout. Based on the variables considered in our study, our results suggest that environmental conditions during early life-history stages are not detrimental for rainbow trout in the study streams. 相似文献
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为了探究三疣梭子蟹受精卵的离体孵化技术及效果,本研究先后开展了受精卵块最适分离液种类及作用条件的筛选、分离液处理不同发育期受精卵离体孵化的差异、分离液处理对受精卵卵膜的结构影响,及分离液处理受精卵对孵化后幼体活力的影响实验。结果显示,木瓜蛋白酶是一种较为理想的分离液,在浓度为0.09 g/mL,分离时间30 min时,分离率可达到99%以上;经过分离液处理后的各期受精卵均能孵化出幼体,卵内溞状幼体期离体胚胎孵化率最高,为89.0%±3.3%,未经处理的对照组为70.0%±4.8%;卵裂期离体胚胎孵化率最低,为58.0%±3.9%,对照组为31.0%±2.3%,与对照组相比,处理组的孵化率明显提高。透射电镜结果显示,经过分离液处理的受精卵卵膜结构疏松,且厚度降低,符合处理组孵化率增加这一现象,干露、福尔马林溶液胁迫和行为学测试对不同处理组的幼体进行质量评价的结果显示,处理组和对照组幼体活力无显著差异。研究表明,实验所获得的分离液可以有效提高三疣梭子蟹受精卵的分离率和孵化率,且不影响幼体质量,可为三疣梭子蟹及其他甲壳动物受精卵的离体孵化提供参考。本研究可为三疣梭子蟹的苗种繁育提供新的技术手段,也将为基因编辑辅助育种等技术的实施奠定基础。 相似文献
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青蛤受精卵和幼虫密度对孵化和生长的影响 总被引:1,自引:1,他引:1
本文报道了青蛤受精卵密度对孵化的影响和青蛤幼虫培养密度对生长的影响。受精卵密度在26个/ml、30个/ml、38个/ml、44个/ml、53个/ml时,孵化率在87.8~89.9%之间;受精卵密度为53个/ml时,幼虫畸形率较高。实验发现,幼虫的培养密度应控制在6~11个/ml,幼虫变态后,减少培养密度,可以提高幼虫成活率。 相似文献
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为了研究盐度对褐点石斑鱼(Epinephelus fuscoguttatus)♀×清水石斑鱼(Epinephelus polyphekadion)♂杂交后代(简称杂交石斑鱼)受精卵孵化、卵黄囊仔鱼形态及活力的影响,将杂交石斑鱼受精卵置于不同盐度水体中,观察盐度胁迫下卵径、油球径、孵化率及畸形率,以及孵化后卵黄囊仔鱼形态变化和活力状况。结果表明,在本研究条件下,受精卵卵径随盐度降低而增大,油球径不随盐度变化而变化。在盐度18~36范围内均能孵化仔鱼,但因盐度不同孵化率和初孵仔鱼畸形率有显著差异(P0.05);随盐度升高,孵化率呈先升后降的变化,初孵仔鱼畸形率则相反。盐度24~36组孵化后3 d仔鱼卵黄囊几乎完全消耗,组间无显著差异(P0.05)。在盐度24~36范围内,随着盐度的增加,仔鱼的不投饵存活系数(SAI值)呈先升后降的变化曲线;盐度30~33范围内,初孵仔鱼SAI值大于5;5日龄仔鱼全部死亡,半数死亡时间出现在孵化后3.5 d。综上分析表明,该杂交石斑鱼受精卵孵化及仔鱼培育的适宜盐度范围为24~36,最适盐度范围为30~33。 相似文献