| 利用shRNA真核表达载体干涉雌性鸡胚性腺中芳香化酶基因(CYP19A1)的表达 |
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| 引用本文： | 朱晓亮,徐桂云,杨宁,李俊英,郑江霞.利用shRNA真核表达载体干涉雌性鸡胚性腺中芳香化酶基因(CYP19A1)的表达[J].农业生物技术学报,2011,19(6). |
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| 作者姓名： | 朱晓亮 徐桂云 杨宁 李俊英 郑江霞 |
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| 作者单位： | 中国农业大学动物科学技术学院,北京,100193 |
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| 基金项目： | 国家自然科学基金资助项目(No.30901017)资助 |
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| 摘 要： | 本研究通过把构建的短发夹结构RNA(shRNA)真核表达载体导入鸡胚,检测鸡胚性腺分化期质粒载体在鸡胚体内代谢情况及雌性鸡胚性腺中芳香化酶基因(CYP19A1)mRNA表达效率,进而探讨利用该方法在鸡胚体内进行特定基因干涉的可行性.实验针对CYP19A1基因构建了4条特异性表达载体,一条非特异性表达载体.每个实验组选取45个新鲜种蛋作为实验材料进行胚盘下腔注射,并设立空白对照组.鸡胚发育12d时,检测各处理组鸡胚肝脏组织中质粒存在情况,并取其左侧性腺组织进行目标基因的荧光定量分析.研究结果表明,导入组在12日胚龄时所有鸡胚基因组中均可检测到绿色荧光蛋白基因(EGFP)；荧光定量结果显示,导入特异性表达载体cyp-580、cyp-1083和cyp-1295后,对应雌性鸡胚性腺CYP19A1 mRNA表达效率显著低于空白对照组,干涉效率分别为:73％、52％和85％；特异性表达载体cyp-1403组CYP19A1mRNA表达效率与空白对照组相比有所降低但无显著性差异.本实验为诱导鸡胚性反转提供了新方法并建立了鸡胚发育期特定基因体内干涉新平台.
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| 关 键 词： | 鸡胚 芳香化酶基因(CYP19A1) RNA干扰(RNAi) 短发夹结构RNA(shRNA) 性反转 |
Interference of Polypeptide 1 Gene(CYP19A1) Expression by shRNA Eukaryotic Expression Vector in Female Chick Embryo Gonads |
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| Zhu Xiaoliang
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Xu Guiyun
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Yang Ning
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Li Junying
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Zheng Jiangxia.Interference of Polypeptide 1 Gene(CYP19A1) Expression by shRNA Eukaryotic Expression Vector in Female Chick Embryo Gonads[J].Journal of Agricultural Biotechnology,2011,19(6). |
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| Authors: | Zhu Xiaoliang Xu Guiyun Yang Ning Li Junying Zheng Jiangxia |
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| Institution: | Zhu Xiaoliang Xu Guiyun Yang Ning Li Junying Zheng Jiangxia* College of Animal Science and Technology,China Agricultural University,Beijing 100193,China |
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| Abstract: | In this study,we have constructed and injected short hairpin RNA(shRNA) eukaryotic expression vectors into fertilized eggs.Upon incubation,the transferred plasmids were analyzed for their metabolism in the embryo gonads during gonadal differentiation,and their interference on the expression of polypeptide 1(CYP19A1) gene(belonging to cytochrome P450,subfamily A,family 19) was assayed subsequently.The feasibility of the method was then further discussed for gene-specific interference in the chick embryo.In t... |
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| Keywords: | Chick embryo Cytochrome P450 family 19 subfamily A polypeptide 1(CYP19A1) RNA interference(RNAi) Short hair RNA(shRNA) Sex reversal |
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