No abstract available.
Aged, 80 and over ; Anti-Bacterial Agents/pharmacology ; Bacterial Proteins/*genetics ; China ; Drug Resistance, Multiple, Bacterial ; Humans ; Male ; Methyltransferases/*genetics ; Microbial Sensitivity Tests ; RNA, Ribosomal, 16S/genetics/metabolism ; Serratia marcescens/drug effects/*enzymology/*genetics/isolation & purification ; Urinary Tract Infections/diagnosis/microbiology

Serratia marcescens jn01 was employed as the host for the isolation of phages from environmental sewage. One strain of phage named SmPjn was purified by picking transparent plaque with 2mm diameter and clear edge on the double-layer agar repeatedly. Electron micrographs indicated that the phage head was icosahedral with head size and tail length of (58 +/- 2.16) x (55 +/- 0.47) nm and (7 +/- 1.25) nm, respectively. On the basis of the morphology, this phage belongs to the family Podoviridae. Host-range determination revealed that the phage was capable of infecting the other two isolates of S. marcescens, P25 and CMCC41002. The optimal multiplicity of infection was 1. A one-step growth curve of SmPjn indicated that the latent period and burst size were estimated at 50 min and 1,125 pfu/cell, respectively . Genomic DNA of SmPjn was above 27kb in size and could be digested by Hind Ill and EcoR I into 11 and 9 visible fragments after electrophoresis, respectively. A novel Podoviridae-phage infecting S. marcescens was firstly reported in China.
China ; DNA, Viral ; genetics ; isolation & purification ; metabolism ; Host Specificity ; Podoviridae ; genetics ; growth & development ; isolation & purification ; Restriction Mapping ; Serratia marcescens ; physiology

To efficiently produce non-specific nuclease (NU) of Serratia marcescens through recombinant overexpression approach and to characterize the purified NU. The nuclease gene was amplified from the genomic DNA of Serratia marcescens by PCR and fused into vector pMAL-c4X with maltose binding protein (MBP) tag. The recombinant vector verified by DNA sequencing was transformed into Escherichia coli BL21. The expressed MBP-NU was purified through the amylose resin and its catalytic characters were analyzed. The results showed the NU gene had 97% identities with the reported S. marcescens nuclease gene and intracellularly expressed in E. coli BL21. The optimal expression conditions were 37 degrees C, 0.75 mmol/L IPTG with 1.5 h induction. The purified MBP-NU exhibited non-specific nuclease activity, able to degrade various nucleic acids, including RNA, single-stranded DNA and double-stranded DNA that was circular or linear. Its optimal temperature was 37 degrees C and optimal pH 8.0. From 1 L culture broth 10.8 mg NU could be purified with a specific activity of 1.11x10(6) U/mg. The catalytic activity of NU was not inhibited by reagents such as EDTA (0.5 mmol/L), PMSF (1 mmol/L) and KCl (150 mmol/L) commonly used in protein purification.
Base Sequence ; Endodeoxyribonucleases ; biosynthesis ; genetics ; Endoribonucleases ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Maltose-Binding Proteins ; genetics ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification ; Serratia marcescens ; enzymology

A minimal test scheme, consisting of deoxyribonuclease (DNase) and tween 80 hydrolysis (TEH) together with a few other biochemical tests, was used to make tentative identification of Serratia marcescens from clinical specimens. The identifications were reevaluated by testing comprehensive biochemical characteristics of 52 isolates, and all were found to be correct. The biochemical reactions of the isolates were very homogenous, showing typical characteristics of the species except in the urease test and acid production from sucrose, adonitol and inositol. These facts support the feasibility of the use of the minimal identification scheme. Pigment production was noted only in 7 isolates invalidating the value of this characteristic for the identification. Fifty-seven isolates were tested for their antibiotic susceptibility. They were found most frequently susceptible to gentamicin (47.4%), chloramphenicol (35.0%) and kanamycin (28.1%). Many isolates (49.1%) were multiply resistant to ampicillin, chloramphenicol, gentamicin, kanamycin, streptomycin and tetracycline.
Antibiotics/pharmacology* ; Cells, Cultured ; Drug Resistance, Microbial ; Human ; Microbial Sensitivity Tests ; Serratia marcescens/drug effects ; Serratia marcescens/isolation & purification ; Serratia marcescens/metabolism*