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采用Matchmaker酵母双杂交系统,将对虾传染性皮下及造血组织坏死病毒(Infection hypodermal and hematopoietic necrosis virus,IHHNV)编码蛋白NS1、NS2和CP序列分别构建到酵母猎物载体p GADT7和诱饵载体p GBKT7上,分别转化至酵母AH109以检测重组猎物载体和诱饵载体的自激活作用及对宿主的毒性作用,发现无自激活作用和毒性作用,随后将重组猎物载体和诱饵载体两两共转至酵母AH109中,涂布于SD/-Leu/-Trp固体培养基上,再点种至SD/-Ade/-His/-Leu/-Trp/X-α-gal固体培养基以鉴定编码蛋白间的相互作用。表型鉴定结果显示,只有共转化有p GADT7-CP/p GBKT7-CP的酵母重组子能够在SD/-Ade/-His/-Leu/-Trp/X-α-gal上生长并显蓝,而其它重组子均不能在其上生长,表明病毒的CP能够自身互作,而其他编码蛋白间无相互作用。为了进一步研究病毒CP自身互作的作用位点,分别从CP的N端和C端截短若干个氨基酸序列,结果发现CP的自身互作是高度敏感的,任何较少氨基酸序列的缺失都将导致其自身互作的丧失。本研究为深入探讨病毒的组装机制和致病机理奠定了理论基础。 相似文献
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从铜绿微囊藻(Microcystis aeruginosa)PCC7820中克隆了生物钟基因kaiB,并将其克隆入酵母双杂交系统的两个质粒pGADT7和pGBKT7中。经测序验证后,将重组质粒pGADT7-kaiB和pGBKT7-kaiB转化酵母菌AH109进行自激活性和自身相互作用的检测和验证。结果表明,铜绿微囊藻生物钟蛋白KaiB无自激活性,KaiB蛋白自身能发生相互作用。因此,可用KaiB蛋白为诱饵利用酵母双杂交系统筛选文库钓取与KaiB发生相互作用的蛋白。 相似文献
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In this paper the optimal precursors for wintertime Eurasian blocking onset are acquired by solving a nonlinear optimization problem whose objective function is constructed based on a blocking index with a triangular T21,three-level,quasi-geostrophic global spectral model.The winter climatological state is chosen as the reference basic state.Numerical results show that the optimal precursors are characterized by a baroclinic pattern with a westward tilt with height,which are mainly located upstream of the blocking region.For an optimization time of 5 days,these perturbations are mainly localized over the Northeast Atlantic Ocean and continental Europe.With the extension of the optimization time to 8 days,these perturbations are distributed more upstream and extensively in the zonal direction.Wave spectrum analysis reveals that the optimal precursors are composed of not only synoptic-scale(wave numbers 5-18) waves,but planetary-scale(wave numbers 0-4) waves as well.The synoptic-scale optimal precursors are mainly located in the mid-latitude area,while the planetary-scale optimal precursors focus primarily on the highlatitude region.The formation of a strong planetary-scale positive blocking anomaly is accompanied by the reinforcement of synoptic-scale perturbations and further fragmentation into two branches,in which the northern branch is generally stronger than the southern one.The eddy forcing arising from the selfinteraction of synoptic-scale disturbances is shown to be crucial in triggering the dipole blocking anomaly,and the planetary-scale optimal precursor provides the initial favorable background conditions for blocking onset. 相似文献
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近岸带三组成波耦合作用的观测与分析 总被引：2，自引：0，他引：2
利用近岸带两个测点的波浪同步观测资料，通过二阶谱分析了三组成波的非线性相互作用。对波谱峰频自相互作用的二阶相干系数b^2(fp，fp）及对应的二阶位相值β(fp，fp)、最大二阶谱幅值B最大(f1，f2)和最大二阶相干系数b^2最大(f1，f2)的组成波频率分析结果表明：(1)弱频散作用的过渡水深区存在着明显的非共振三组成波耦合作用；(2)包括谱峰频在内的自相互作用是近岸区波耦合作用的重要形式；(3)随着水深减小，趋近碎波带，非线性耦合作用进一步增强，谱能量以向低频转移为主变为向高、低频两侧转移，波形趋于前倾。 相似文献
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