Avtor/Urednik		Stres, Blaž
Naslov		Molekulsko odkrivanje in sledenje bakterij rodu Pseudomonas in podskupine P. stutzeri v vzorcih iz okolja
Prevedeni naslov		Molecular detection and monitoring of the genus Pseudomonas and subgroup P. stutzeri in environmental samples
Tip		monografija
Kraj izdaje		Ljubljana
Založnik		Medicinska fakulteta
Leto izdaje		2001
Obseg		str. 87
Jezik		slo
Abstrakt		Specificity of existing primers for the specific detection of the genus Pseudomonas and P. stutzeri subgroup was evaluated in computer based analysis of 16S rRNA genes. New combinations of existing primers and new regions were used for preparation of optimized PCR. Specificity was experimentally determined using genomic DNA of 48 cultures belonging to the genus Pseudomonas and 10 belonging to P.stutzeri subgroup. Eleven wild type strains were isolated from soil and identified using PCR as members of the genus, eight as P. stutzeri and identified with commercial kit API 20 NE. Optimized PCR was used to develop competitive PCR (c-PCR). PCR products were analyzed using Bioanalyzer Agilent 2100 and the results obtained by regression equation. c-PCR for the specific detection of members of the genus Pseudomonas enabled us detection of 3,2 x 10 cells per gram of soil added. Methods of DNA isolation and purification used in this work provided too few target sequences to allow us specific enumeration of members of P. stutzeri subgroup with developed c-PCR. In our future work we will try to improve methods for DNA isolation and purification from soil and optimize developed c-PCR methods.
Deskriptorji		PSEUDOMONAS SOIL MICROBIOLOGY RNA, RIBOSOMAL, 16S POLYMERASE CHAIN REACTION DNA PROBES OLIGONUCLEOTIDE PROBES DNA, BACTERIAL