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The three types of reconstructed embryos(reconstructed embryo of goat-rabbit, goat-bovine and goat-goat) were produced respectively by nuclear transfer using goat ear firbroblast cells as donors and rabbit, bovine and goat ooctyes as recipients, and the factors of cytoplast environments influencing cloned embryo development in vitro, cytoplast effect in nuclear transfer, the methods of transferring cloned embryo and the effects of pregnancy factor on implantation of cloned embryo were studied in this experiment, in order to supply basis for solving the difficulty of implantation of inter-species cloned embryo in recipient and improving the efficiency of cloning.
本试验以波尔山羊耳成纤维细胞为供体，分别以牛、羊、兔的卵母细胞为受体进行体细胞核移植，构建了山羊-兔、山羊-牛和山羊-山羊三种重构胚。研究了不同的胞质环境对重构胚体外发育的影响与核移植中的胞质效应，并对异种克隆胚胎采用了去透明带移植和配种后再移植的方法，探讨移植方法和妊娠因子对异种克隆胚胎在受体动物体内发育的影响，旨在为解决异种克隆胚移植不易着床发育的难题和为提高动物克隆效率提供理论依据。
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Gondii, were cloned by PCR respectively. The PCR products were digested by the corresponding enzymes and ligatd into the intermedial vectors. Finally, the inducible RNAi vector, pBSK-HSP70/5UTR-IntronC-HSP70/3UTR, with the HSP70 gene promotor as a promotor, the intron C sequence ofβ-tubulin gene as intervening sequence, 3UTR sequences of HSP70 gene as transcription stop signals, was constructed successfully, and the results of sequencing were correct. 3The construction of the inherited and inducible RNAi vector system of T. gondii: The fragment of SAG1/5UTR-eGFP-SAG1/3UTR in pBSK-SAG1/GFP vector was cloned into the vector of pBSK-HSP70/5UTR-IntronC-HSP70/3UTR to construct pBSK-GFP-Hairpin vector, then the fragment of GFP-Hairpin in pBSK-GFP-Hairpin vector was cloned into pHANA-0.5 vector.
弓形虫可诱导的反向重复序列RNAi载体的构建：设计引物，通过PCR分别扩增弓形虫HSP70基因5&UTR启动子序列(HSP70／5UTR)、HSP70基因3&UTR序列(HSP70／3UTR)及β-微管蛋白基因内含子C序列，通过酶切连接，构建以弓形虫热休克蛋白HSP70基因启动子进行驱动的，以β-微管蛋白基因内含子C序列作为间隔序列，以HSP70基因3UTR序列作为转录终止信号的反向重复序列RNAi载体pBSK-HSP70／5UTR-IntronC-HSP70／3UTR，序列测定结果正确。
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CDNA fragments of mnh6 and mtp1 genes were cloned from M. grisea appressorium cDNA library and full length DNA fragments of these genes were cloned from genomic DNA by LD-PCR.
利用长距离PCR从稻瘟病菌附着胞cDNA文库中克隆了mnh6和mtp1基因的cDNA序列，并从基因组DNA中克隆了mnh6和mtp1基因的全长DNA序列。
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CLONED：克隆结束
CLON ING 开始克隆 | CLONED 克隆结束 | DEACT IVATED Agent 已休眠
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cloned gene：克隆化基因
PCR 扩增产物的快速克隆] | cloned gene 克隆化基因 | cloning 克隆,克隆化
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cloned gene：选殖基因
Clone contig 殖株连续体 209 | Cloned gene 选殖基因 434 | Cluster analysis 丛集分析 215
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Cloned Node：克隆节点
Clock Circuit 时钟电路 | Cloned Node 克隆节点 | COM 组件对象模型
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cloned calf：克隆牛
beef scare 疯牛病 | 34、cloned calf 克隆牛 | 35、Mandarin putonghua 普通话
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