Comparison of mycoplasma ovipneumoniae isolates using bacterial restriction endonuclease DNA analysis and SDS-PAGE

SUMMARYNinety-six isolates of presumptive or confirmedListeria monocytogeneswere obtained from local clinical (30 isolates) or food laboratories (66 isolates). Minimal biochemical analysis identified only 80% of these isolates asL. monocytogenesthe remaining includedL. seeligeri, 1%, or the non-haemolyticL. innocua, 19%. The 27 clinical and 50 food isolates, mainly from meat products, frozen confectionaries, and cheeses, confirmed asL. monocytogeneswere compared biochemically and serologically. Twenty-one isolates, including some strains ofL. innocuaandL. seeligeri, were examined for pathogenicity in immunocompromized mice and 44 typed using bacterial restriction endonuclease DNA analysis (BRENDA). Only isolates ofL. monocytogeneswere found to be pathogenic. Biovar-typing of the isolates was unreliable and provided poor discrimination. Serogroups 1/2 and 4 predominated among clinical and food isolates and BRENDA provided better discrimination among isolates. Ten stable and reproducible restriction patterns were observed among theListeriasp. isolates studied. Overall, a combination of techniques gave the best discrimination and indicated their potential for use as epidemiological tools.

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Polypeptide and antigenic variability among strains of Mycoplasma ovipneumoniae demonstrated by SDS-PAGE and immunoblotting

Veterinary Microbiology ◽

10.1016/0378-1135(90)90035-t ◽

1990 ◽

Vol 21 (3) ◽

pp. 241-254 ◽

Cited By ~ 14

Author(s):

D. Thirkell ◽

R.K. Spooner ◽

G.E. Jones ◽

W.C. Russell

Keyword(s):

Antigenic Variability ◽

Sds Page ◽

Mycoplasma Ovipneumoniae

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A Frameshift Mutation in the Human Fibrinogen Aα-chain Gene (Aα [499] Ala Frameshift Stop) Leading to Dysfibrinogen San Giovanni Rotondo

Thrombosis and Haemostasis ◽

10.1055/s-0037-1616752 ◽

2001 ◽

Vol 86 (12) ◽

pp. 1483-1488 ◽

Cited By ~ 12

Author(s):

Gennaro Vecchione ◽

Rosa Santacroce ◽

Francesca D’Angelo ◽

Bruno Casetta ◽

Maria Luisa Papa ◽

...

Keyword(s):

Mass Spectrum ◽

Dna Analysis ◽

Stop Codon ◽

Premature Stop Codon ◽

Disulfide Bridge ◽

Chain Gene ◽

Human Fibrinogen ◽

Single Nucleotide ◽

Sds Page

SummaryWe have investigated a 53-yr-old asymptomatic white man with decreased functional, but not immunologic, fibrinogen plasma levels together with prolonged thrombin and reptilase times, detected through routine coagulation studies prior to a surgical procedure. A new heterozygous single nucleotide deletion (C) at position Ala499 within the Aα-chain gene was identified, which predicted changes of the corresponding aminoacids encoded by the subsequent portion of the exon V and the appearance of a premature stop codon at position 518 (A [499]Ala frameshift stop). The new dysfunctional fibrinogen, San Giovanni Rotondo variant, was confirmed in vivo by SDS-PAGE analysis of HPLC-purified fibrinogen chains. Mass spectrum examination of the abnormal HPLC-purified peak gave an estimated mass (56,088 Da) similar to that predicted by DNA analysis of the mutated Aα-chain gene (56,088 Da) and, after tryptic digestion, the truncated Aα-chain was shown only in the propositus, who also carried normal Aα-chain. In addition, mass spectrum analysis of the tryptic digest of the abnormal chain confirmed the presence of a new and unpaired cysteine at the last position that was predicted to form a disulfide bridge with human serum albumin. Immuno-blot analysis confirmed that fibrinogen San Giovanni Rotondo variant, but not normal fibrinogen, contained substantial amounts of albumin. Present findings confirm that truncated Aα-chain lacking part of the terminal domain may be incorporated into mature fibrinogen molecules and normally secreted in the bloodstream.

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Restriction endonuclease DNA analysis of Clostridium difficile.

Journal of Clinical Microbiology ◽

10.1128/jcm.25.12.2402-2404.1987 ◽

1987 ◽

Vol 25 (12) ◽

pp. 2402-2404 ◽

Cited By ~ 40

Author(s):

B W Wren ◽

S Tabaqchali

Keyword(s):

Clostridium Difficile ◽

Restriction Endonuclease ◽

Dna Analysis

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Fibrinogen Milano XII: a dysfunctional variant containing 2 amino acid substitutions, Aα R16C and γ G165R

Blood ◽

10.1182/blood.v98.2.351 ◽

2001 ◽

Vol 98 (2) ◽

pp. 351-357 ◽

Cited By ~ 7

Author(s):

Bettina Bolliger-Stucki ◽

Susan T. Lord ◽

Miha Furlan

Keyword(s):

Dna Analysis ◽

Ethylenediaminetetraacetic Acid ◽

Degradation Products ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Structural Defects ◽

Thrombin Time ◽

Sds Page ◽

Α Helix ◽

Immunologic Assays

Fibrinogen Milano XII was detected in an asymptomatic Italian woman, whose routine coagulation test results revealed a prolonged thrombin time. Fibrinogen levels in functional assays were considerably lower than levels in immunologic assays. Polymerization of purified fibrinogen was strongly impaired in the presence of calcium or ethylenediaminetetraacetic acid (EDTA). Two heterozygous structural defects were detected by DNA analysis: Aα R16C and γ G165R. As seen previously with other heterozygous Aα R16C variants, thrombin-catalyzed release of fibrinopeptide A was 50% of normal. Additionally, the release of fibrinopeptide B was delayed. Immunoblotting analysis with antibodies to human serum albumin indicated that albumin is bound to Aα 16 C. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) analysis of plasmin digests of fibrinogen Milano XII in the presence of calcium or EDTA showed both normal and novel D1 and D3 fragments. Further digestion of abnormal D3 fragments by chymotrypsin resulted in degradation products of the same size as the fragments derived from normal fibrinogen. SDS-PAGE analysis under reducing conditions showed no difference between normal fibrinogen and fibrinogen Milano XII or between their plasmic fragments. Circular dichroism analysis revealed a shift in the mean residual ellipticity and a significant reduction of the α-helix content in the variant D3 fragment. It is concluded that the Aα-chain substitution is mainly responsible for the coagulation abnormalities, whereas the substitution in the γ-chain induced a conformational change in the D3 fragment.

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Serratia entomophilabacteriophages: host range determination and preliminary characterization

Canadian Journal of Microbiology ◽

10.1139/m97-152 ◽

1997 ◽

Vol 43 (11) ◽

pp. 1069-1073 ◽

Cited By ~ 6

Author(s):

Maureen O'Callaghan ◽

Trevor A. Jackson ◽

Travis R. Glare

Keyword(s):

Host Range ◽

Restriction Endonuclease ◽

Large Scale ◽

Dna Analysis ◽

Bacterial Pathogen ◽

Restriction Endonuclease Analysis ◽

Scale Production ◽

Bacterial Isolates ◽

Host Range Determination ◽

Range Determination

Eight bacteriophages specific to Serratia entomophila, a commercially available bacterial pathogen of the New Zealand grass grub (Costelytra zealandica), were characterized by host range determination, morphology and restriction endonuclease patterns of DNA. Phages were originally isolated from grass grub larvae and fermenter broth where phages had disrupted large-scale production of S. entomophila. Seven of the phages (CW1–CW5, BC, and BT) had heads similar in size (approximately 60 × 60 nm) and long noncontractile tails (185 × 10 nm). Phage AgRP8 (P8) had a smaller head and a short tail structure. Restriction endonuclease analysis divided the phages into four groups: CW2, CW4, CW5, BC, and BT gave identical patterns, while CW1, CW3, and P8 each gave different patterns. Six distinct phage groups were distinguished by host range determination, after screening phages against 70 bacterial isolates: CW1, CW2/CW4, CW3, CW5, BC/BT, and P8. While confirming the indicated groupings by DNA analysis, it was possible to distinguish between some of the phages in the largest group: CW2/4 could be distinguished from CW5 and BC/BT. Screening of soil bacterial isolates of S. entomophila against nondiluted phages will aid in monitoring the establishment and persistence of strains applied for biological control of the grass grub.Key words: Serratia entomophila, bacteriophage, morphology, phage typing, host range.

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