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| 角膜混浊小鼠突变候选基因Map3k1克隆与序列分析 | |
| 引用本文： | 吴刘成,刘春,蒋荧梅,王生存,邵义祥.角膜混浊小鼠突变候选基因Map3k1克隆与序列分析[J].动物医学进展,2011,32(4):1-5. |
| 作者姓名： | 吴刘成 刘春 蒋荧梅 王生存 邵义祥 |
| 作者单位： | 1. 南通大学实验动物中心,江苏南通,226001 2. 南通大学比较医学研究所,江苏南通,226001 3. 南通大学实验动物中心,江苏南通226001;南通大学比较医学研究所,江苏南通226001 |
| 基金项目： | 国家自然科学基金，江苏省自然科学基金，南通大学自然科学基金，南通大学研究生科技创新计划项目 |
| 摘 要： | 对具有角膜混浊表型的B6-Co突变系小鼠突变候选基因Map3k1进行克隆及测序分析,寻找该突变系小鼠Map3kl基因的突变位点.以小鼠Map3k1基因的mRNA、全长及上游5 kb序列设计引物,分别以基因组DNA和mRNA为模板,采用PCR和RT-PCR技术分段扩增目的基因,将目的片段连接在T栽体上,转化至感受态细胞,... |
| 关 键 词： | B6-Co突变系小鼠 Map3k1基因 PCR扩增 序列分析 |
Cloning and Sequence Analysis of Mutant Candidate Gene of Map3k1 in Cornea Opacity Mouse |
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| WU Liu-cheng,LIU Chun,JIANG Ying-mei,WANG Sheng-cun,SHAO Yi-xiang.Cloning and Sequence Analysis of Mutant Candidate Gene of Map3k1 in Cornea Opacity Mouse[J].Progress In Veterinary Medicine,2011,32(4):1-5. | |
| Authors: | WU Liu-cheng LIU Chun JIANG Ying-mei WANG Sheng-cun SHAO Yi-xiang |
| Institution: | 1,2(1.Laboratory Animals Centre,Nantong University,Nantong,Jiangsu,226001,China;2.Institute of Comparative Medicine,Nantong University,Nantong,Jiangsu,226001,China) |
| Abstract: | To find mutational site of Map3k1 gene,clone and sequene the Map3k1 gene in B6-Co mutant strain mouse with cornea opacity phenotype,The primers were designed according to mRNA,full length sequence,and upstream about 5 kb sequence of mouse Map3k1 gene.The target gene was amplified by PCR and RT-PCR in which the PCR templates were genomic DNA and mRNA,and then recombinant vectors were transformed to comptent cells after DNA fragments were connected to the T vector.The positive clones were selected and plasmid DNA extraction was detected by electophoresis.The fragments by EcoRⅠdigestion for identification of positive clones and at the end sequencing in Sangon Biotech(Shanghai) Co.,Ltd.20 exons of Map3k1 and its upstream regulatory sequence were amplified.Sequencing results indicated the T base in sequence of Map3k1 chromosome 13 112 559 574 was replaced by A in B6-Co mouse by sequece alignment with genome data base,with coding protein 314 Leu replacement by Glu.It was expected to be investigated ON the mechanism of the gene expression regulation and the protein clipping. |
| Keywords: | B6-Co mutant strain mouse Map3k1 gene PCR amplification sequence analysis |
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