Obtaining Of A Molecular Marker Of A Functional Bacterial Strain And It's Use In The Waste Water Treating System

The objectives of this study are to develop RAPD markers of functional bacterial strains FA-2 isolated from the fatty chemical waste water treating system, and convert them into SCAR markers for detecting the functional bacterial strains. A total of 40 arbitrary 10-mer oligonucleotide primers were screened on the genomic DNA of two functional bacterial strains (named FA-2, FAS) and the control (coded 1. 150$). Among the PCR products, a 700 bp specific DNA fragment of FA-2 was obtained and cloned into pMD 18-T Vector. After sequencing of the fragment, a pairs of SCAR primers ranging from 18 to 25 bases in length were designed according to the nucleotide sequence. Based on the results of different factors, PCR conditions with the pairs of specific SCAR markers were set up . After optimizing PCR conditions, we converted the RAPD marker into SCAR marker by PCR amplifying. In order to verify the specificity of the SCAR marker, we used the pairs of primers to amplify 20 culture samples. The result shows that the SCAR marker is reliable, and could be used as a molecular marker to detect the functional bacterial strain of FA-2 in the waste water treating system.In order to establish a kind of rapid and efficacious molecular method to quantificationally detect the functional bacterial strains in the waste water treating system, we must prepare total DNA from treating system sludge entirely. Based on different methods and results,a rapid method for high yield DNA extracting from the sludge was established. Electrophoretic and PCR results showed that the quantity of the DNA prepared from sludge was good enough for PCR amplifying. Combined with FQ-PCR (fluorescence quantitive polymerase chain reaction), we can establish a rapid method to detect the functional bacterial strains quantificationally in the waste water treating system.