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Myostatin基因打靶载体的构建及猪胎儿成纤维细胞Myostatin基因的敲除 总被引:1,自引:0,他引:1
目的:用缺口修复等技术构建Myostatin(肌肉生长抑制素,MSTN)基因打靶载体,并对大白猪胎儿成纤维体细胞进行转染,获得基因敲除细胞。方法与结果:首先构建用于MSTN基因同源长臂(LA)的抓捕载体,然后在大肠杆菌内利用Red同源重组系统介导的缺口修复,从含大白猪MSTN基因座的细菌人工染色体上亚克隆9.9 kb的LA到抓捕载体上,经过部分序列测定,同源性为100%;通过PCR获得1.4 kb的同源短臂(SA);将LA和SA连入载体pLOXP,构建含有neo和tk正负筛选标记基因的MSTN基因打靶载体pLOXP-MSTN-KO;将线性化的pLOXP-MSTN-KO通过电转染整合到大白猪胎儿成纤维细胞基因组中,利用G418和丙氧鸟苷进行药物筛选,获得抗性细胞克隆890个,通过PCR和DNA测序鉴定获得基因敲除的细胞克隆4个。结论:构建了有效的MSTN基因打靶载体,通过转染获得基因敲除细胞,为利用体细胞核移植制备MSTN基因敲除猪奠定了基础。 相似文献
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突触传递的长时程抑制(long-term depression,LTD)和长时程增强(long term-potentiation,LTP)是突触可塑性的两种重要形式,并且与学习记忆密切相关。本文探讨Sprague-Dawley(SD)大鼠在海马齿状回区(dentate gyrus,DG)注射36h孵育形成的寡聚体Aβ1-4230d后,在体海马前穿通纤维-齿状回通路(perforant path-dentate gyrus pathway,PP-DG)的突触可塑性和空间记忆能力的变化。2.5月龄SD大鼠随机分为寡聚体Aβ1-42注射组[即阿尔茨海默病(Alzheimer’s disease,AD)模型组,n=12]和正常对照组(n=12),分别在双侧海马DG区注射5μg寡聚体Aβ1-42或生理盐水。应用Morris水迷宫检测大鼠空间记忆能力。同时运用神经电生理在体胞外记录技术,检测寡聚体Aβ1-42引起的海马双脉冲易化(paired pulse facilitation,PPF)、LTD、LTP等突触可塑性形式的变化。结果显示:(1)AD模型组大鼠空间记忆能力下降(P<0.05);(2)寡聚体Aβ1-42降低... 相似文献
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Yu-Jie Liu Ze-Kun Yang Jiong-Ju Hao Bao-Liang Ma Yun Zhang Hong-Wei Yang 《Plasmonics (Norwell, Mass.)》2018,13(4):1201-1207
In this paper, modified transmission matrix method is used to construct one-dimensional multilayer composite membrane structure doped with graphene defect. The optimal construction can be found to realize reciprocity transmission by comparing the influence of the time inversion-symmetry and space inversion-symmetry doped on nonreciprocal transmission. The simulation results show that it cannot ensure the nonreciprocal transmission with rotatory material only. Nonreciprocal transmission should be designed through the structure damage of the space inversion-symmetry to realize it. The structure absorption peak position will move to the direction of the wavelength increase along with the increase of thickness of rotation media. The structure shows the approximate perfect absorption characteristics. 相似文献
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记述了我国蚤蝇科蚤蝇属Phora 1新种,又突蚤蝇Phora furcularis Liu et Wang,sp.nov.和1新纪录种,缺齿蚤蝇P.edentata Schmitz.P.furcularis与P.zherikhini Mostovski相似,但前者额向后渐狭,生殖背板侧叶无钩状突,且左侧尾叶后背部生有"Y"形脊.模式标本存于沈阳大学生物与环境工程学院. 相似文献