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 polym...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.展开更多
The intestinal Oxalobacter Formigenes were isolated in 30 cases of urolithiasis and in 45 controls. The biologic characters and morphology of the bacteria were also observed. The results showed that the colony counts ...The intestinal Oxalobacter Formigenes were isolated in 30 cases of urolithiasis and in 45 controls. The biologic characters and morphology of the bacteria were also observed. The results showed that the colony counts in urolith group 9 (mean 103/g. faeces) were significantly less than that of controls (mean 108/g. faeces) (P<0. 001). It is believed that the lesser amount of oxalobacter formigenes in urolith was the important factor of the calcium oxalate calculi formation.展开更多
基金This project was supported by a grant from National Natural Sciences Foundation of China (No. 30371423)
文摘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.
文摘The intestinal Oxalobacter Formigenes were isolated in 30 cases of urolithiasis and in 45 controls. The biologic characters and morphology of the bacteria were also observed. The results showed that the colony counts in urolith group 9 (mean 103/g. faeces) were significantly less than that of controls (mean 108/g. faeces) (P<0. 001). It is believed that the lesser amount of oxalobacter formigenes in urolith was the important factor of the calcium oxalate calculi formation.
