A PCR technique based on the Hip1 interspersed repetitive sequence distinguishes cyanobacterial species and strains.
The use of primers based on the Hip1 sequence as a typing technique for cyanobacteria has been investigated. The discovery of short repetitive sequence structures in bacterial DNA during the last decade has led to the development of PCR-based methods for typing, i.e., distinguishing and identifying, bacterial species and strains. An octameric palindromic sequence known as Hip1 has been shown to be present in the chromosomal DNA of many species of cyanobacteria as a highly repetitious interspersed sequence. PCR primers were constructed that extended the Hip1 sequence at the 3' end by two bases. Five of the 16 possible extended primers were tested. Each of the five primers produced a different set of products when used to prime PCR from cyanobacterial genomic DNA. Each primer produced a distinct set of products for each of the 15 cyanobacterial species tested. The ability of Hip1-based PCR to resolve taxonomic differences was assessed by analysis of independent isolates of Anabaena flos-aquae and Nostoc ellipsosporum obtained from the CCAP (Culture Collection of Algae and Protozoa, IFE, Cumbria, UK). A PCR-based RFLP analysis of products amplified from the 23S-16S rDNA intergenic region was used to characterize the isolates and to compare with the Hip1 typing data. The RFLP and Hip1 typing yielded similar results and both techniques were able to distinguish different strains. On the basis of these results it is suggested that the Hip1 PCR technique may assist in distinguishing cyanobacterial species and strains.
Smith
J K
JK
IENS, Division of Biological Sciences, Lancaster UniversityBailrigg, Lancaster LA1 4YQUK.
Parry
J D
JD
IENS, Division of Biological Sciences, Lancaster UniversityBailrigg, Lancaster LA1 4YQUK.
Day
J G
JG
Culture Collection of Algae and Protozoa (CCAP), IFE, Windermere LaboratoryThe Ferry House, Far Sawrey, Ambleside, Cumbria LA22 OLPUK.
Smith
R J
RJ
IENS, Division of Biological Sciences, Lancaster UniversityBailrigg, Lancaster LA1 4YQUK.
D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D002412:Cations; D000458:Cyanobacteria / Q000145:classification* / Q000235:genetics; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D004755:Enterobacteriaceae / Q000145:classification / Q000235:genetics; D020071:Interspersed Repetitive Sequences / Q000235:genetics*; D008274:Magnesium; D016133:Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*; D012680:Sensitivity and Specificity; D013045:Species Specificity; D013291:Streptococcus / Q000145:classification / Q000235:genetics; D019914:Taq Polymerase; D013696:Temperature; D013698:Templates, Genetic
D001426:Bacterial Proteins; D002412:Cations; D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D004268:DNA-Binding Proteins; D019914:Taq Polymerase; D008274:Magnesium