| 拟蜘蛛拖丝蛋白基因在小鼠颗粒细胞中的整合及其鉴定 |
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| 引用本文： | 张滕子,王春生,李晶,宁方勇,朴善花,安铁洙.拟蜘蛛拖丝蛋白基因在小鼠颗粒细胞中的整合及其鉴定[J].黑龙江畜牧兽医,2011(7). |
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| 作者姓名： | 张滕子 王春生 李晶 宁方勇 朴善花 安铁洙 |
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| 作者单位： | 东北林业大学生命科学学院;东北农业大学动物科技学院; |
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| 基金项目： | 国家自然科学基金项目(30771538); 东北林业大学引进人才科研启动基金项目 |
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| 摘 要： | 为了探讨拟蜘蛛拖丝蛋白基因二聚体(2S)在小鼠颗粒细胞基因组中整合和建立转拟蜘蛛拖丝蛋白基因的小鼠颗粒细胞系的可能性,试验将在前期工作中人工合成的拟蜘蛛拖丝蛋白基因2S连接到具有CMV启动子、IRES序列和GFP报告基因的表达载体,并转染小鼠颗粒细胞。结果表明:原代培养的幼鼠卵巢的颗粒细胞在培养后的第4天达到85%汇合,继代培养1~7代时每代达到85%的汇合时间约为3 d,细胞的形态与原代培养细胞相似,呈现多角形或梭形;筛选阳性细胞的最佳G418浓度为500μg/mL;将线性化的CMV-2S-pIRES2-EGFP转染颗粒细胞,再用500μg/mLG418筛选12 d后仍有存活细胞,但是续培养细胞的增殖能力逐渐丧失;经PCR鉴定,阳性细胞扩增出目的条带。
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| 关 键 词： | 拟蜘蛛拖丝蛋白基因 小鼠颗粒细胞 整合 鉴定 |
Integration and identification of mouse granular cells with the spider dragline silk protein gene |
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| ZHANG Teng-zi,WANG Chun-sheng,LI Jing,NING Fang-yong,PIAO Shan-hua,AN Tie-zhu.Integration and identification of mouse granular cells with the spider dragline silk protein gene[J].Heilongjiang Animal Science And veterinary Medicine,2011(7). |
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| Authors: | ZHANG Teng-zi WANG Chun-sheng LI Jing NING Fang-yong PIAO Shan-hua AN Tie-zhu |
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| Affiliation: | ZHANG Teng-zi1,WANG Chun-sheng1,LI Jing1,NING Fang-yong1,2,PIAO Shan-hua1,AN Tie-zhu1(1.College of Life Science,Northeast Forestry University,Harbin 150040,China,2.College of Animal Science & Technology,Northeast Agricultural University,Harbin 150030,China) |
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| Abstract: | To discuss the feasibility of establishment of mouse granular cell line with the transgene of spider dragline silk protein gene,in this experiment artificially synthesized spider dragline silk protein gene was linked to the pIRES2-GFP vector with CMV promoter,IRES sequence and GFP report gene,and granular cell was transfected with the recombination plasmid.The results showed that natal mouse primary granular cell spent 4 d to congregate 90% confluence,and cells spent 3 d to congregate 85% confluence in subc... |
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| Keywords: | Spiders dragline silk protein gene mouse granular cells integration identification |
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