On the Role of Restriction Enzymes of Haemophilus in Transformation and Transfection

Gromkova, Rosa; Goodgal, Sol H.

doi:10.1007/978-1-4684-2133-0_19

Cited by 14 publications

(13 citation statements)

References 9 publications

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“…Transfection, at least in the Rd strains studied, is thus sensitive to restriction. This confirms earlier reports on H. influenzae (7,8,30) and on Bacillus subtilis (39). However, when isogenic r-and r+ defectively lysogenic strain Rd recipients were exposed to the homologous DNAs, prophage transformation, as well as antibiotic resistance transformation, was only slightly reduced, at most.…”

supporting

confidence: 93%

Restriction enzymes do not play a significant role in Haemophilus homospecific or heterospecific transformation

Stuy¹

1976

J Bacteriol

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Competent Haemophilus influenzae Rd recipients, either as phage HP1 restricting (r+) or nonrestricting (r-) nonlysogens or defective lysogens, were exposed to deoxyribonucleic acids from various wild-type phage HP1 lysogenic H. influenzae serotype strains (non-encapsulated derivatives of serotypes a, b, c, d, and e), to DNA from lysogenic Haemophilus parahaemolyticus, and to DNA from modified and nonmodified phage HP1. Transformation of antibiotic resistance markers and of prophage markers in homospecific crosses was observed to be unaffected by the recipient restriction phenotype, whereas the transfection response was much reduced in r+ recipients. Heterospecific transformation of prophage markers was reduced by only 80 to 90%, whereas antibiotic resistance marker transformation was 1,000 to 10,000 times lower. Heterospecific transfection was at least 100 times lower than homospecific transfection in both r+ and rrecipients. The general conclusion is that neither class I nor class II restriction enzymes affect significantly the transformation efficiency in homospecific and heterospecific crosses. The efficiency of heterospecific transformation may depend mainly on the deoxyribonucleic acid homology in the genetic marker region.-A general feature of heterospecific bacterial genetic transformation is the low efficiency of marker transfer from donor to heterospecific recipient (for a general review, see reference 1; for Haemophilus, see references 3, 16, 18, 27). Exceptions have been observed, however (14). The extent of the reduction is particular for a given marker. Explanations of this phenomenon can be based on either (i) reduced uptake of the heterospecific deoxyribonucleic acid (DNA), (ii) the presence of so-called restriction enzymes (32) in the recipient that destroy the genetic integrity of adsorbed nonmodified DNA either before or after integration, or (iii) the lack of homology between donor and recipient DNA. Although competent Haemophilus influenzae Rd cells show a reduced uptake of unrelated foreign DNA (15, 28), the uptake of heterologous Haemophilus parainfluenzae DNA appears to be normal (18,34). Moreover, the molecular fate of this DNA is comparable to that of homologous DNA after uptake (3,18,34,36). This appears to rule out a major role of restriction enzymes at the preintegration stage. It has also been shown that H. parainfluenzae DNA is integrated into the H. influenzae genome (18,34). This integration causes the death of a considerable fraction of the recipient cells through induction of the defective prophage (29) that is present in the Rd strain used most widely (4, 33). Since a presumably cured strain (2, 4) is insensitive to this lethal effect and since heterospecific transformation is also normally inefficient in this strain (29), the prophage induction phenomenon is not believed to be the explanation for the reduced heterospecific transformation efficiency. The lowered marker transfer is thus probably caused by events that follow integration (for a thorough discussion, see reference 17...

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supporting

confidence: 93%

Restriction enzymes do not play a significant role in Haemophilus homospecific or heterospecific transformation

Stuy¹

1976

J Bacteriol

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“…3) of the SV40 DNA fragments produced by the H. influenzae restriction endonuclease by analyzing incompletely digested DNA fragments, as well as an overlapping set of fragments, produced by a restriction endonuclease from H. parainfluenzae (ref. 13; Sack and Nathans, manuscript in preparation); these experiments will be reported elsewhere (Danna, Sack, and Nathans, manuscript in preparation). In Fig.…”

Section: Resultsmentioning

confidence: 91%

Bidirectional Replication of Simian Virus 40 DNA

Danna

Nathans

1972

Proc. Natl. Acad. Sci. U.S.A.

215

113

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“…injluenzae type b to be transformed is probably not due to the effect of restriction enzymes. Although Haemophilus bacteria are known to be a rich source of such enzymes, previous studies have demonstrated that they do not play a role in the exclusion of bacterial transforming DNA in uiuo (Gromkova & Goodgal, 1977). In addition, isogenic DNA is usually protected against the action of restriction enzymes.…”

Section: Discussionmentioning

confidence: 99%

Genetic Transformation in Encapsulated Clinical Isolates of Haemophilus influenzae Type b

Rowji

Gromkova²,

Koornhof³

1989

Microbiology

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Haemophilus inzuenzae type b strains isolated from children with meningitis, septicaemia and pharyngitis were studied for their ability to undergo genetic transformation by two chromosomal markers, streptomycin resistance and nalidixic acid resistance. Fifty-eight percent of the strains were non-transformable while the remaining 42% showed considerable strain variation with regard to their transformation frequencies, which ranged from 8 x to 1 xThe effect of type b capsule on competence development and transformation activity was studied by comparing encapsulated strains with their non-encapsulated variants. Type b capsule did not inhibit either competence development or transforming efficiency. The lack of transformability in the majority of strains was not due to the presence of a capsule.

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On the Role of Restriction Enzymes of Haemophilus in Transformation and Transfection

Cited by 14 publications

References 9 publications

Restriction enzymes do not play a significant role in Haemophilus homospecific or heterospecific transformation

Restriction enzymes do not play a significant role in Haemophilus homospecific or heterospecific transformation

Bidirectional Replication of Simian Virus 40 DNA

Genetic Transformation in Encapsulated Clinical Isolates of Haemophilus influenzae Type b

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