| 改良红鲫Dmc1基因编码阅读框的克隆及表达 |
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| 引用本文： | 周曼,冯浩. 改良红鲫Dmc1基因编码阅读框的克隆及表达[J]. 激光生物学报, 2012, 0(5): 429-433,445 |
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| 作者姓名： | 周曼 冯浩 |
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| 作者单位： | 湖南师范大学生命科学学院,湖南长沙410081 |
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| 基金项目： | 湖南省杰出青年科学基金（12JJ1005）;教育部新世纪优秀人才支持计划（NCET-11-0971）;教育部留学回国人员科研启动基金（2011-1568-1） |
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| 摘 要： | Dmc1基因是一个在减数分裂前期Ⅰ表达的基因,其表达产物为减数分裂时同源染色体配对所必需。本实验根据普通红鲫Dmc1基因编码阅读框(ORF)序列设计引物,克隆了改良红鲫Dmc1基因(命名为IRCC-Dmc1)的ORF序列并将之插入到cDNA5-FRT/TO载体中,构建了改良红鲫Dmc1基因表达载体FRT/TO-IRCC-Dmc1-Myc。以FRT/TO-IRCC-Dmc1-Myc质粒转染HEK293T细胞,蛋白印迹杂交实验检测到改良红鲫Dmc1基因在HEK293T细胞中有着正确的表达。本文为将来进一步研究改良红鲫Dmc1基因的功能打下了基础。
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| 关 键 词： | Dmc1 改良红鲫 克隆 |
Molecular Cloning and Expression of the Open Reading Frame of Dmcl Gene for Improved Red Crucian Carp |
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| ZHO U Man,FENG Hao. Molecular Cloning and Expression of the Open Reading Frame of Dmcl Gene for Improved Red Crucian Carp[J]. ACTA Laser Biology Sinica, 2012, 0(5): 429-433,445 |
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| Authors: | ZHO U Man FENG Hao |
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| Affiliation: | ( College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China) |
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| Abstract: | Dmcl （disrupted meiotic cDNA）gene is specifically expressed in the premeiotic prophase I during miosis. In this study, we obtained the open reading frame （ORF） of Dmcl gene for IRCC （named IRCC-Dmel ） through PCR ampli- fication, using primers based on the ORF of norulal Red Crucian Carp. The IRCC-Dmcl was inserted into cDNAS- FRT/TO and the recombinant plasmid FRT/TO-IRCC-Dmcl-Myc was constructed. HEK293T cells were transfected with FRT/TO-IRCC-Dmcl -Myc through calcium phosphate method. The data of western blot demonstrated that IRCC-Dmel has strong expression in HEK293T cells. Our study has established a solid foundation for the functional study of IRCC-Dmcl gene |
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| Keywords: | Dmcl Improved Red Crucian Carp cloning |
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