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[Objective] The aim of this study is to understand the effects of donor cell type,embryo stage,number and transfer position on the efficiency of goat transgenic clone.[Method] Using somatic cell nuclear transfer technology,the single goat fetal fibroblasts(GFF)and mammary gland epithelial cells(GMGE)harboring human lactoferrin(hLF)gene were transferred to the enucleated oocyte.Reconstructed karyoplast-cytoplast couplets were fused,activated,and cultured in vitro.Embryos at 2-8 cell stage were transferred into oviduct of synchronized recipients,and blastocysts were transferred into uterine horn.[Result] The pregnancy rate was similar between GFF and GMGE(oviduct transfer:26.47% vs.20.00%),and between oviduct transfer and uterine horn transfer(26.47% vs.25.00%)for GFF group;pregnancy rate in the group with the mean number of embryo transferred per recipient of 21.2 was significantly higher than in those the 5.93 group and 9.64 group(40.00% vs.26.67% and 21.43%).[Conclusion] These results indicate that pregnancy rate of goat transgenic clone couldn’t be affected by donor cell type,embryo stage and transfer position but be done by the number of embryo transferred per recipient.In addition,the study also suggests the feasibility of making transgenic goat using GMGE as donor cells.  相似文献   
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[目的]探讨供体细胞类型、移植胚胎发育阶段、数量及部位对山羊转基因克隆效率的影响。[方法]利用体细胞核移植技术将转染人乳铁蛋白基因hLF的山羊胎儿成纤维细胞(GFF)和乳腺上皮细胞(GMGE)移植到MII期去核卵母细胞内,经电融合、激活、体外培养后,2~8细胞期克隆胚被移植到同期发情山羊的输卵管内,囊胚被移植到子宫角内。[结果]GFF与GMGE的妊娠率相近(输卵管移植妊娠率分别为26.47%及20.00%);在GFF,输卵管移植的妊娠率与子宫角内移植妊娠率接近(分别为26.47%及25.00%),输卵管移植胚胎平均数为21.2组的妊娠率显著高于5.93组和9.64组(40.00%及26.67%,21.43%)。[结论]供体细胞类型、移植胚胎的发育阶段及移植部位对山羊转基因克隆效率的影响不大,但对于输卵管移植,受体羊移植胚胎数量对妊娠率有明显的影响。此外,该研究还提示了利用成年羊乳腺上皮细胞制作转基因动物的可行性。  相似文献   
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构建真核表达载体pEGFP-C1-hLYZ,并从基因水平研究人溶菌酶基因在体外培养的牛乳腺上皮细胞中的表达,为核移植提供转基因供体细胞.采用PCR的方法从人溶菌酶质粒中扩增854 bp人溶菌酶基因(包括完整的编码框446 bp).并克隆到pMD18-T载体上,用限制性内切酶Hind Ⅲ和BamHⅠ双酶切.将人溶菌酶基因的酶切片段(883 bp)定向克隆到pEGFP-C1的多克隆位点中,构建重组表达质粒pEGFP-hLYZ,并进行Hind Ⅲ和Bam HI双酶切鉴定.转染体外培养的牛乳腺上皮细胞,48 h后观察其表达情况.PCR检测溶菌酶基因的表达.成功构建了人溶菌酶真核表达载体,观察到表达绿色荧光蛋白的牛乳腺上皮细胞,并检测到溶菌酶基因.  相似文献   
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水通道蛋白与植物的抗旱性   总被引:12,自引:0,他引:12  
水通道蛋白(aquaporin,AQP)构成水分运输的特异性通道,促进水的长距离运输,调节细胞内外水分平衡,并运输其它小分子物质。有专一性、高效性、可调控性和活性差异性等特点。本文就近几年来AQP的结构、功能、调节机理及其与植物抗旱性研究进展作一简要综述。  相似文献   
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