Cloning and Identification of frc Gene from Oxalobacter Frmigenes  

作　　者：孔德波 陈志强 叶章群 杨为民 姚林方 郭辉 刘冠琳 曾令启 

机构地区：[1]Department of Urology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology

出　　处：《Journal of Huazhong University of Science and Technology(Medical Sciences)》2007年第2期190-192,共3页华中科技大学学报（医学英德文版）

基　　金：This project was supported by a grant from National Natural Sciences Foundation of China (No. 30371423)

摘　　要：The cloning and identification offrc gene from Oxalobacterformigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacterformigenes was extracted, frc gene fragment was amplified by polymerase chain reaction （PCR） and linked with pEGFP-CI. 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 fro 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 thatfrc gene cloned from the Oxalobacterformigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.The cloning and identification offrc gene from Oxalobacterformigenes in the intestines of Chinese people were conducted. The genomic DNA of Oxalobacterformigenes was extracted, frc gene fragment was amplified by polymerase chain reaction （PCR） and linked with pEGFP-CI. 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 fro 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 thatfrc gene cloned from the Oxalobacterformigenes in the intestines of Chinese people can be expressed in eucaryotic 293 cells and keep its enzyme activity.

关 键 词：oxalobacter FRC UROLITHIASIS gene cloning 

分 类 号：R691[医药卫生—泌尿科学]