| Cloning and Identification of frc Gene from Oxalobacter Frmigenes |
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| 作者姓名： | 孔德波 陈志强 叶章群 杨为民 姚林方 郭辉 刘冠琳 曾令启 |
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| 作者单位： | Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Department of Urology Tongji Hospital Tongji Medical College Huazhong University of Science and Technology，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China，Wuhan 430030 China |
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| 摘 要： | The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chi- nese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.
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| 关 键 词： | 产甲酸草酸杆菌 frc基因 基因克隆 鉴定 尿石形成 |
| 收稿时间： | 2006-09-09 |
Cloning and identification of <Emphasis Type="Italic">frc</Emphasis> gene from <Emphasis Type="Italic">Oxalobacter frmigenes</Emphasis> |
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| Kong Debo,Chen Zhiqiang,Ye Zhangqun,Yang Weimin,Yao Linfang,Guo Hui,Liu Guanlin,Zeng Lingqi.Cloning and Identification of frc Gene from Oxalobacter Frmigenes[J].Journal of Zuazhong University of Science and Technology: Medical Edition,2007,27(2):190-192. |
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| Authors: | Kong Debo Chen Zhiqiang Ye Zhangqun Yang Weimin Yao Linfang Guo Hui Liu Guanlin Zeng Lingqi |
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| Institution: | Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030,China |
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| Abstract: | The cloning and identification of frc gene from Oxalobacter formigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacter formigenes was extracted. frc gene fragment was amplified by polymerase chain reaction (PCR) and linked with pEGFP-C1. The recombinant plasmid was designated pEGFP-frc and was identified by restriction-enzyme digestion and sequencing. Human embryo kidney 293 cells were transfected with pEGFP-frc, then RT-PCR and Western blotting were performed to detect the expression of frc gene. The length of frc gene was found to be 1287 bp, and the homology of nucleotides and amino-acid residue with the sequence in GenBank was 95.88% and 99.07%. Bright green fluorescent light could be observed in 293 cells transfected with the pEGFP-frc. frc mRNA and fusion protein FCoAT-EGFP were detected in the cells. It is concluded that frc gene cloned from the Oxalobacter formigenes in the intestines of Chi- nese people can be expressed in eucaryotic 293 cells and keep its enzyme activity. |
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| Keywords: | oxalobacter frc urolithiasis gene cloningKONG Debo male born in 1978 Candidate for Doctoral Degree |
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