scholarly journals Improved gfp and inaZ Broad-Host-Range Promoter-Probe Vectors

2000 ◽  
Vol 13 (11) ◽  
pp. 1243-1250 ◽  
Author(s):  
William G. Miller ◽  
Johan H. J. Leveau ◽  
Steven E. Lindow
Keyword(s):  
Host Range ◽  
Reporter Gene ◽  
Half Life ◽  
The Other ◽  
Broad Host Range ◽  
Promoter Probe ◽  
Restriction Sites ◽  
Unique Restriction ◽  
Selection For ◽  
Derivatives Of

A new set of broad-host-range promoter-probe vectors has been constructed. One subset contains the pVS1 and p15a replicons and confers resistance to either gentamicin or kanamycin. The other set contains the broad-host-range replicon from pBBR1 and confers resistance to kanamycin, tetracycline, ampicillin, or spectinomycin/streptomycin. Both plasmid sets are highly stable and are maintained without selection for more than 30 generations in several bacterial taxa. Each plasmid contains a promoter-probe cassette that consists of a multicloning site, containing several unique restriction sites, and gfp or inaZ as a reporter gene. The cassette is bound by transcriptional terminators to permit the insertion of strong promoters and to insulate the cassette from external transcription enabling the detection of weak or moderate promoters. The vector suite was augmented with derivatives of the kanamycin-resistant gfp promoter-probe plasmids that encode Gfp variants with different half-life times.

Construction of a broad-host-range pneumococcal promoter-probe plasmid

Gene ◽  
1990 ◽  
Vol 90 (1) ◽  
pp. 163-167 ◽  
Author(s):  
Eduardo Diaz ◽  
JoséL. Garcia
Keyword(s):  
Host Range ◽  
Broad Host Range ◽  
Promoter Probe

Isolation and genetic analysis of Azospirillum brasilense Nif− mutants

1983 ◽  
Vol 29 (8) ◽  
pp. 968-972 ◽  
Author(s):  
P. Jara ◽  
B. Quiviger ◽  
P. Laurent ◽  
C. Elmerich
Keyword(s):  
Genetic Analysis ◽  
Host Range ◽  
Azospirillum Brasilense ◽  
Nitrogenase Activity ◽  
Plasmid Vector ◽  
Ecori Fragment ◽  
Broad Host Range ◽  
Broad Host Range Plasmid ◽  
Ethylmethane Sulfonate ◽  
Derivatives Of

After ethylmethane sulfonate mutagenesis of Azospirillum brasilense strain 7000, mutants devoid of nitrogenase activity were isolated. Partial diploids were constructed by introducing plasmids pAB35 and pAB36 into the Nif− mutants. The two plasmids were derivatives of the broad host-range plasmid vector pRK290. Plasmid pAB35 contained a 6.7 kilobase pairs (kb) EcoRI fragment which carried the nifHDK gene cluster cloned from strain 7000. Plasmid pAB36 contained the same fragment from which a 2.6-kb PstI fragment that likely covers nifK, and a part of nifD was deleted. The restoration of a Nif+ phenotype by pAB35, but not by pAB36, was observed in the case of mutant 7571, which might be impaired in a structural gene for the nitrogenase complex.

Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes

Gene ◽  
1995 ◽  
Vol 166 (1) ◽  
pp. 175-176 ◽  
Author(s):  
Michael E. Kovach ◽  
Philip H. Elzer ◽  
D. Steven Hill ◽  
Gregory T. Robertson ◽  
Michael A. Farris ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Host Range ◽  
Cloning Vector ◽  
Broad Host Range ◽  
Derivatives Of

Host-specific effects of the korA-korB operon and oriT region on the maintenance of miniplasmid derivatives of broad host-range plasmid RK2

Plasmid ◽  
1989 ◽  
Vol 21 (2) ◽  
pp. 99-112 ◽  
Author(s):  
Thomas J. Schmidhauser ◽  
David H. Bechhofer ◽  
David H. Figurski ◽  
Donald R. Helinski
Keyword(s):  
Host Range ◽  
Broad Host Range ◽  
Specific Effects ◽  
Broad Host Range Plasmid ◽  
Derivatives Of ◽  
Plasmid Rk2

scholarly journals New broad-host-range promoter probe vectors based on the plasmid RK2 replicon

2001 ◽  
Vol 195 (1) ◽  
pp. 91-96 ◽  
Author(s):  
Pedro Miguel Santos ◽  
Ilaria Bartolo ◽  
Janet Martha Blatny ◽  
Elisabetta Zennaro ◽  
Svein Valla
Keyword(s):  
Host Range ◽  
Broad Host Range ◽  
Promoter Probe ◽  
Plasmid Rk2

New mini-Tn5 derivatives for insertion mutagenesis and genetic engineering in Gram-negative bacteria

1995 ◽  
Vol 41 (11) ◽  
pp. 1053-1055 ◽  
Author(s):  
M. F. Alexeyev ◽  
I. N. Shokolenko ◽  
T. P. Croughan
Keyword(s):  
Genetic Engineering ◽  
Gene Cloning ◽  
Insertion Mutagenesis ◽  
Broad Host Range ◽  
Gram Negative Bacteria ◽  
Origin Of Replication ◽  
Gram Negative ◽  
Broad Host Range Plasmid ◽  
Restriction Sites ◽  
Unique Restriction

Five mini-Tn5 derivatives encoding resistance to Km, Cm, Gm, Tc, and Sm, coupled with the polylinker of the pBluescriptII plasmid, were constructed. These derivatives are carried by an ampicillin-resistant plasmid that has a conditional origin of replication from plasmid R6K and origin of conjugal transfer from the broad host range plasmid RP4. The new vectors are smaller than those previously described and possess numerous unique restriction sites inside the minitransposons for gene cloning in addition to SfiI and NotI sites found in their predecessors.Key words: R6K γ-origin, suicide vectors, RP4 oriT, chromosomal insertion.

Broad-host-range IncP-4 plasmid R1162: effects of deletions and insertions on plasmid maintenance and host range

1982 ◽  
Vol 152 (1) ◽  
pp. 140-150
Author(s):  
R Meyer ◽  
R Laux ◽  
G Boch ◽  
M Hinds ◽  
R Bayly ◽  
...  
Keyword(s):  
Host Range ◽  
Hot Spot ◽  
Broad Host Range ◽  
Plasmid Maintenance ◽  
Cis Acting ◽  
E Coli ◽  
Sequence Organization ◽  
Restriction Sites ◽  
Recessive Defect

R1162 is an 8.7-kilobase (kb) broad-host-range replicon encoding resistance to streptomycin and sulfa drugs. In vitro deletion of 1.8-kb DNA between coordinates 3.0 and 5.3 kb did not affect plasmid maintenance, but a Tn1 insertion at coordinate 6.3 kb led to a recessive defect in plasmid maintenance. The only cis-acting region necessary for plasmid replication appears to lie between the Tn1 insertion at coordinate 6.3 kb and a second Tn1 insertion at coordinate 6.5 kb. All R1162 sequences between position 6.5 kb and the EcoRI site at coordinate 8.7/0 kb were dispensible for replication in Escherichia coli and Pseudomonas putida. Plasmids carrying insertions in a variety of restriction sites in an R1162::Tn1 derivative were unstable in P. putida but stable in E. coli. Tn5 insertions in R1162 showed a hot spot at coordinate 7.5 kb. A Tn5 insertion at coordinate 8.2 kb appeared to mark the 3' end of the streptomycin phosphotransferase coding sequence. All R1162::Tn5 derivatives showed specific instability in Pseudomonas strains but not in E. coli. The instability could be relieved by internal deletions of Tn5 sequences. In the haloaromatic-degrading Pseudomonas sp. strain B13, introduction of an unstable R1162::Tn5 plasmid led to loss of ability to utilize m-chlorobenzoate as a growth substrate. Our results showed that alteration of plasmid sequence organization in nonessential regions can result in restriction of plasmid host range.

Enhancement of Nucleopolyhedrovirus Activity in Helicoverpa zea (Boddie) and Pseudoplusia includens (Walker) Larvae with a Fluorescent Brightener

2001 ◽  
Vol 36 (2) ◽  
pp. 162-168 ◽  
Author(s):  
S. Y. Young
Keyword(s):  
Host Range ◽  
Helicoverpa Zea ◽  
Heliothis Armigera ◽  
Autographa Californica ◽  
The Other ◽  
Pseudoplusia Includens ◽  
Broad Host Range ◽  
Fluorescent Brightener ◽  
Occlusion Bodies ◽  
Anagrapha Falcifera

The enhancement of nucleopolyhedrovirus (NPV) activity by Tinopal® LPW (Tinopal), a stilbene fluorescent brightener, was compared in Helicoverpa zea (Boddie) and Pseudoplusia includens (Walker) using an on-diet bioassay method. Enhancement of the homologous NPV of each species was compared with three heterologous NPV that have a broad host range. In H. zea, the LC50 of H. zea NPV (HzSNPV) alone was 128 occlusion bodies (OBs)/cup, and the LC50 of it and Heliothis armigera NPV (HaMNPV) did not differ significantly. The activity of both viruses improved 18.6 fold when the OB suspension contained 1.0% Tinopal. The LC50s of Autographa californica NPV (AcMNPV) and Anagrapha falcifera NPV (AfMNPV) without Tinopal in H. zea were greater than that of HzSNPV. However, the increase in activity of AcMNPV and AfMNPV at the highest concentrations of Tinopal was two to three fold greater than the increase in activity of HzSNPV. The LC50 of P. includens NPV (PiSNPV) (856 OBs/cup) alone in P. includens was similar to that of AfMNPV and AcMNPV, and much less than that of HaMNPV (19,947 OBs/cup). The addition of Tinopal to the treatment suspension of all four viruses resulted in significantly lower LC50s at all Tinopal concentrations in P. includens. The highest concentration of Tinopal (1.0%) in the OB suspension lowered the LC50 of PiSNPV by 142.7 and AfMNPV by 89.7 fold. Tinopal in the OB suspension lowered the LC50 of AcMNPV and HaMNPV, but they remained less effective than PiSNPV with Tinopal. HaMNPV at all concentrations of Tinopal was much less active in P. includens than the other viruses with or without Tinopal.

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