Isolation and characterization of two temperate phages from Lactococcus lactis ssp. cremoris C3

1992 ◽  
Vol 38 (5) ◽  
pp. 398-404 ◽  
Author(s):  
Audrey W. Jarvis ◽  
Vaughan R. Parker ◽  
Moana B. Bianchin
Keyword(s):  
Lactococcus Lactis ◽  
Dna Hybridization ◽  
Base Plate ◽  
Restriction Endonuclease Analysis ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Isolation And Characterization ◽  
Dna Fragment ◽  
Endonuclease Analysis

Lactococcus lactis ssp. cremoris C3 was shown to contain two prophages, C3-T1 and C3-T2, both inducible bymitomycin C. The phages were morphologically indistinguishable, having an isometric head (55 nm), a noncontractile tail (142 nm), and a distinctive base plate. The phages C3-T1 and C3-T2 were differentiated from one another by restriction endonuclease analysis of their DNA, and genome sizes of 37.8 (C3-T1) and 32.5 kb (C3-T2). Southern blot DNA hybridization suggested a maximum homology between the phage genomes of 50%. Phage C3-T1 was propagated and the phage obtained was designated C3-TIℓ. Phage C3-T2 was not propagated despite tests with a large number of possible indicator strains. Restriction enzyme analysis of phage C3-T1ℓ genome yielded a 37.8-kb circular restriction map. The packaging site (pac) and site of integration into the bacterial chromosome (att) were located and an EcoRI DNA fragment containing the att site was cloned. Only one C3-T1 att site was present in the C3 chromosome. No homology was detected between DNA from C3-T1ℓ and DNA from lytic phages commonly isolated from cheese wheys. Key words: lactococcal phages, temperate phages, lysogeny, evolution.

scholarly journals Restriction Endonuclease Analysis Discriminates Bordetella bronchiseptica Isolates

2000 ◽  
Vol 38 (12) ◽  
pp. 4387-4393 ◽  
Author(s):  
Randy E. Sacco ◽  
Karen B. Register ◽  
Gwen E. Nordholm
Keyword(s):  
Restriction Enzyme ◽  
Host Species ◽  
Restriction Endonuclease Analysis ◽  
Bordetella Bronchiseptica ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Dna Fragments ◽  
Chromosomal Dna ◽  
Agarose Gels ◽  
Endonuclease Analysis

One hundred ninety-five Bordetella bronchisepticaisolates from 12 different host species worldwide were characterized by restriction enzyme analysis (REA). These isolates had previously been categorized into 19 PvuII ribotypes. Twenty restriction endonucleases were evaluated for use in REA. Digestion of chromosomal DNA with HinfI, followed by submarine electrophoresis in agarose gels and staining with ethidium bromide, produced DNA fragments in the 4.0- to 10-kb range, which readily discriminated B. bronchiseptica isolates, resulting in 48 fingerprint patterns. Moreover, AluI digestion of chromosomal DNA produced 39 distinct fingerprint profiles with DNA fragments ranging from 6.0 to 20.0 kb. While REA frequently provided more discriminatory power than ribotyping, there were examples where the use of ribotyping was more discriminatory than REA. Passage of selected isolates up to passage 25 did not change the REA profile. Moreover, the Bvg phase did not alter the fingerprint profile of chromosomal DNA from B. bronchiseptica strains digested with HinfI orAluI. Based on the results presented herein, the combination of REA and ribotyping should provide valuable information in understanding the molecular epidemiology of B. bronchiseptica infections.

scholarly journals Characterization of Dutch porcine Serpulina (Treponema) isolates by restriction endonuclease analysis and DNA hybridization

1992 ◽  
Vol 138 (9) ◽  
pp. 1929-1934 ◽  
Author(s):  
A. A. H. M. T. Huurne ◽  
M. Van Houten ◽  
M. B. H. Koopman ◽  
B. A. M. Van Der Zeijst ◽  
W. Gaastra
Keyword(s):  
Restriction Endonuclease ◽  
Dna Hybridization ◽  
Restriction Endonuclease Analysis ◽  
Endonuclease Analysis

Isolation and characterization of antibiotic-resistance plasmids in thermophilic bacilli

1985 ◽  
Vol 31 (4) ◽  
pp. 339-345 ◽  
Author(s):  
Takayuki Hoshino ◽  
Takayuki Ikeda ◽  
Hiroyuki Narushima ◽  
Noboru Tomizuka
Keyword(s):  
Antibiotic Resistance ◽  
Southern Hybridization ◽  
Tetracycline Resistance ◽  
Restriction Endonuclease Analysis ◽  
Kanamycin Resistance ◽  
Isolation And Characterization ◽  
Thermophilic Bacilli ◽  
Resistance Plasmids ◽  
Endonuclease Analysis

Four antibiotic-resistance plasmids isolated from thermophilic bacilli were characterized in detail. Three tetracycline-resistance (Tcr) plasmids were designated as pTHT9 (7.7 kilobases (kb)), pTHT15 (4.5 kb) and pTHT22 (8.4 kb). From the results of restriction endonuclease analysis and the subsequent Southern hybridization, these were found to possess extensive genetic homology in the regions that include the replication origin and the Tcr gene. Detailed restriction maps of the smallest Tcr plasmid pTHT15 and a kanamycin-resistance (Kmr) plasmid pTHN1 (4.8 kb) were constructed. The positions of antibiotic-resistance loci and regions essential for plasmid replication were determined by cloning plasmid fragments in Bacillus subtilis. These four plasmids were found to replicate and express the resistance genes stably in both B. subtilis and B. stearothermophilus.

scholarly journals Paratuberculosis in Farmed Deer: Case Reports and DNA Characterization of Isolates of Mycobacterium Paratuberculosis

1993 ◽  
Vol 5 (4) ◽  
pp. 567-571 ◽  
Author(s):  
Geoffrey W. de Lisle ◽  
Gary F. Yates ◽  
Desmond M. Collins
Keyword(s):  
New Zealand ◽  
Dna Hybridization ◽  
Case Reports ◽  
Restriction Endonuclease Analysis ◽  
Mycobacterium Paratuberculosis ◽  
Dna Characterization ◽  
Pathologic Features ◽  
Histologic Changes ◽  
Endonuclease Analysis

Mycobacterium paratuberculosis was isolated from farmed deer in New Zealand on 21 occasions over a 7-year period. The number of cases increased during the last 3 years of the study. Clinical paratuberculosis was observed in 5 deer, 3 other cases came from grossly normal animals that reacted to a tuberculin skin test, and the remaining 13 animals had tuberculous lesions that were identified at meat inspection. Pathologic features of the lesions in these 13 cases included necrosis and mineralization in lymph nodes draining the gastrointestinal tract. The histologic changes in these lesions were very similar to those caused by Mycobacterium bovis and members of the M. avium complex. Characterization of 20 of the isolates of M. paratuberculosis by restriction endonuclease analysis and DNA hybridization revealed that 3 of these isolates were identical to New Zealand sheep isolates and 17 were the same as cattle isolates. The source of the cervine infections was not determined.

Characterization of Trypanosoma cruzi isolates from Paraguay, using restriction enzyme analysis of kinetoplast DNA

1992 ◽  
Vol 86 (3) ◽  
pp. 231-237 ◽  
Author(s):  
T. Mimori ◽  
M. Maldonado ◽  
M. Samudio ◽  
A. Rojas De Arias ◽  
R. Moreno ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Restriction Enzyme ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Kinetoplast Dna

scholarly journals Characterization of two BHV-4 strains isolated in the Czech Republic

2010 ◽  
Vol 55 (No. 3) ◽  
pp. 106-112 ◽  
Author(s):  
V. Fichtelova ◽  
K. Kovarcik
Keyword(s):  
Reference Strain ◽  
Fragment Size ◽  
Blot Hybridization ◽  
Size Variation ◽  
Southern Blot Hybridization ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
The Czech Republic ◽  
Herpesvirus 4

This study describes the isolation of bovine herpesvirus 4 (BHV-4) from the respiratory tract of animals suffering from respiratory disease. DNA of new isolates, CH and Ni, was cleaved with <I>Bam</I>HI, <I>Eco</I>RI and <I>Hind</I>III in restriction enzyme analysis and the fragments were identified by co-migration with the restriction profile of the reference strain Movar 33/63 cleaved with the appropriate endonuclease. Typical profiles with polyrepetitive DNA (prDNA) fragments were detected. In order to localize the size variation within the obtained digestion fragments, Southern blot hybridization was performed. Differences between the isolates CH, Ni were localized in both the prDNAs and the unique central part of the genome and were restricted to fragment size variation.

scholarly journals Genetic and structural characterization of an amino acid dimorphism in glycoprotein Ib alpha involved in platelet transfusion refractoriness

Blood ◽  
1992 ◽  
Vol 79 (11) ◽  
pp. 3086-3090 ◽  
Author(s):  
M Murata ◽  
K Furihata ◽  
F Ishida ◽  
SR Russell ◽  
J Ware ◽  
...  
Keyword(s):  
Platelet Transfusion ◽  
Platelet Reactivity ◽  
Recombinant Expression ◽  
Structural Basis ◽  
Enzyme Analysis ◽  
Restriction Enzyme Analysis ◽  
Definitive Evidence ◽  
Naturally Occurring ◽  
Alpha Gene

Abstract The platelet-specific alloantigen, Siba, located within the alpha- subunit of the glycoprotein (GP) Ib-IX membrane receptor, has been found to be involved in the pathogenesis of platelet transfusion refractoriness. We have identified the existence of a naturally occurring threonine/methionine dimorphism at position 145 of the GPIb alpha sequence, and determined that the Siba antigen corresponds to the molecule containing methionine145. The diallelic codons can be detected by restriction enzyme analysis of amplified genomic DNA fragments from the GPIb alpha gene. Evaluation of 61 healthy blood donors showed that the allele frequencies are 89% and 11% for the threonine145 and methionine145 codons, respectively. A positive correlation exists between platelet reactivity with the anti-Siba antibody and the presence of a methionine145-encoding allele. Moreover, recombinant expression of two soluble GPIb alpha fragments differing only at residue 145, provided definitive evidence that the human anti-Siba antibody reacts only with the molecule containing methionine145. These results explain the structural basis of the Siba human alloantigen system and define screening methodologies useful in transfusion medicine to match donor and recipient platelets accordingly.

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