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Characterisation of the replication and maintenance of a bacillus subtilis plasmid

Andrew Peter Gleave

Characterisation of the replication and maintenance of a bacillus subtilis plasmid

by Andrew Peter Gleave

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Published by University of Birmingham in Birmingham .
Written in English


Edition Notes

Thesis (Ph.D)-University of Birmingham, Dept of Genetics, 1989.

Statementby Andrew Peter Gleave.
ID Numbers
Open LibraryOL13890964M

In this paper we describe the Isolation and characterization of single strand origins (SSOs) of several cryptic Bacillus subtilis plasmids which use the rolling-circle mechanism of replication. Alonso JC, Viret JF, Tailor RH. Plasmid maintenance in Bacillus subtilis recombination-deficient mutants. Mol Gen Genet. Jun; ()– Canosi U, Morelli G, Trautner TA. The relationship between molecular structure and transformation efficiency of some S. aureus plasmids isolated from B. subtilis. Mol Gen Genet.

We determined the effect of various Bacillus subtilis dna(Ts) mutations on pSM non-permissive temperature, plasmid replication depends on the dnaB, dnaC, dnaG and dnaI initiation replication products, but does not require the dnaA elongation is performed by DNA polymerase III, since replication is blocked at non-permissive temperature in dna(Ts) mutants. Abstract pAMβ1 is a plasmid isolated from Enterococcus faecalis which replicates in Bacillus subtilis by a unidirectional theta mechanism. It has been shown previously that initiation of pAMβ1.

The gram-positive bacterium Bacillus megaterium has several advantages over other recombinant protein production hosts. In contrast to Bacillus subtilis, B. megaterium does not possess alkaline proteases and is known for the stable replication and maintenance of plasmids ().The bacterium readily secretes proteins into the growth medium. For this study, the commercially interesting levansucrase. Canosi U, Iglesias A, Trautner TA. Plasmid transformation in Bacillus subtilis: effects of insertion of Bacillus subtilis DNA into plasmid pC Mol Gen Genet. ; (4)– Chestukhin AV, Shemyakin MF, Kalinina NA, Prozorov AA. Some properties of ATP dependent deoxyribonucleases from normal and rec-mutant strains of Bacillus subtilis.


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Characterisation of the replication and maintenance of a bacillus subtilis plasmid by Andrew Peter Gleave Download PDF EPUB FB2

A kb fragment of the large (approximately 55 kb) Bacillus subtilis plasmid pLS20 containing all the information for autonomous replication was cloned and sequenced. In contrast to the parental plasmid, derived minireplicons were unstably by: Characterization of the replication region of the Bacillus subtilis plasmid pLS a novel type of replicon Wilfried J.J.

Meijer, Arjo L. de Boer, Sandra van Tongeren, Gerard Venema and Sierd Bron Published in: Nucleic Acids Research () 23; A kb fragment of the large (approximately 55 kb) Bacillus subtilis plasmid pLS20 containing all the information for autonomous replication was cloned and sequenced.

In contrast to the parental plasmid, derived minireplicons were unstably maintained. Characterization ofthereplication region ofthe plasmid pLS20 containing all the information for replication. orfA shows homology to the is chromosomal genes rapA (spoOL, gsiA). involved in plasmid maintenance.

In addition, the knowledge obtained has been used used for the construction of improved cloning vectors for is. Since segregational plasmid stability is closely associated with plasmid replication, a considerable part of this thesis deals with studies on plasmid replication in is.

The plasmid pBC16 ( kbases), originally isolated from Bacillus cereus, determines tetracycline resistance and can be transformed into competent cells of B. subtilis. A miniplasmid of pBC16 (pBC), 2,7 kb) which has lost an EcoRI fragment of pBC16 retains the replication functions and the tetracycline resistance.

This plasmid which carries only one EcoRI site has been joined in vitro to. An isogenic set of 11 recombination-deficient mutant strains of Bacillus subtilis has been constructed. Whereas plasmid pUB is stably maintained in such Rec- cells, the high copy number plasmid pC is unstable.

Instability in Rec- strains could be mostly attributed to the deleterious effect of the presence of the plasmid on the Rec- cells' growth capability. Plasmid pL32 from the Natto strain of Bacillus subtilis belongs to a group of low-copy-number plasmids in gram-positive bacteria that replicate via a theta mechanism of replication.

DISCUSSION The replication functions of two resident plasmids of B. thuringiensis, pHT and pHT, were used to construct derivatives able to replicate in B. subtilis. The presence of the transposon Tn and their thermosensitive replication confers an additional interest to these plasmids in view of genetic studies in gram-positive bacteria.

A series of hybrid plasmids consisting of pC or pUB and B. subtilis DNA were constructed. In contrast to plasmid pC, purified monomeric forms of such plasmids were active in transformation, provided the recipient cells were recombination proficient.

Similarly the monomers of PC derived plasmids, containing bacteriophage ϕ DNA were able to transform ϕ lysogenic but not.

Plasmid pL32 from the Natto strain of Bacillus subtilis belongs to a group of low-copy-number plasmids in gram-positive bacteria that replicate via a theta mechanism of replication. We studied the DNA region encoding the replication protein, RepN, of pLS32, and obtained the following by: We determined the effect of various Bacillus subtilis dna(Ts) mutations on pSM replication.

At non-permissive temperature, plasmid replication depends on the dnaB, dnaC, dnaG and dnaI initiation replication products, but does not require the dnaA function. Replication of the Plasmids pBOE, pBOE, andpBOE in B.

subtilis All three plasmids were maintained at a very low copy number (inferred from the yields of plasmid DNA in whole-cell lysates) in B. subtilis compared to pHV and pHV (the entire replicon ofpC ) (Fig.

2), and all the plasmids were segregationally unstable with loss. Restriction enzyme analysis, cloning, and sequencing showed that large (more than 90 kb) plasmids isolated from different Bacillus subtilisstrains are identical in structure of the region ensuring stable inheritance of plasmid replicons and are widespread in Belarussian environmental strains of B.

subtilis. Characterization of the minimal origin required for replication of the streptococcai plasmid plP in Bacillus subtilis S. Brantl^ and D. Behnke Institute for Moiecuiar Biology, Beutenbergstr Jena, Germany. Summary By using deletional analysis the origin of replication, oriR, of the streptococcai plasmid plP in Bacillus.

The effects of an intercalating dye, ethidium bromide (EtBr), on the initiation of chromosome replication in Bacillus subtilis were studied. Spores of a thymine requiring mutant acquired the ability to initiate one round of replication in the absence of RNA and protein synthesis (initiation potential) during germination in a thymine starved medium.

The full text of this article hosted at is unavailable due to technical difficulties. All of sso in B. pumilus plasmids isolated so far belong to the ssoT 2 subfamily, suggesting that the sso DNA in the B. pumilus plasmids are more homologous than those in the B.

subtilis plasmids. Download: Download full-size image; Fig. Comparison of Single strand DNA (ssDNA) among Bacillus pumilus and Bacillus subtilis plasmids. The introduction into the plasmid of a hairpin structure, whose stem carries a DnaA box, mediates conversion of SS(c) into dsDNA and makes plasmid replication independent of the B.

subtilis dnaB. Chapter IX SUMMARY We have sequenced and analyzed a kb fragment of the 55 kb endogenous Bacillus subtilis plasmid pLS20 containing its replication functions. Just outside the region required for autonomous replication, a segment of 18 bp was identified as being almost identical to part of the major is chromosomal replication.

During early stationary phase, Bacillus subtilis NAFM5 produces capsular poly(γ-glutamic acid) (γPGA, 2× Da), which contains d- and l-glutamate, and then degrades it during late stationary phase. The γ-glutamyltransferase (EC ; GGT) of this strain successively hydrolysed γPGA from the amino-terminal end, to yield both d- and l-glutamate.is.

phosphatase before the ligation step. The Cloning of the SSO of the cryptic is plasmid pTA Next, pWM was used to select the SSO of the cryptic plasmid pTA of is IAM (Uozumi et al., ).

This RCM plasmid was chosen because it is maintained stably in spite of its low copy-number and, therefore, is.Vol.No. 5 Characterization ofPBSX, a Defective Prophage ofBacillus subtilis HEATHERE. WOOD,*MICHAELT. DAWSON,ELL Department ofGenetics, Trinity College, Dublin2, Ireland Received 28 November/Accepted 7 February PBSX, adefective Bacillus subtilis prophage, mapsto the metA-metCregion ofthe chromosome.

DNA(33 .