A broad-host-range in vivo pop-out and amplification system for generating large quantities of 50-to 100-kb genomic fragments for direct DNA sequencing

Gene ◽  
1996 ◽  
Vol 179 (1) ◽  
pp. 181-188 ◽  
Author(s):  
Jadwiga Wild ◽  
Zdenka Hradečná ◽  
György Pósfai ◽  
Waclaw Szybalski
2003 ◽  
Vol 77 (3) ◽  
pp. 1964-1976 ◽  
Author(s):  
Alexej Prassolov ◽  
Heinz Hohenberg ◽  
Tatyana Kalinina ◽  
Carola Schneider ◽  
Lucyna Cova ◽  
...  

ABSTRACT All hepadnaviruses known so far have a very limited host range, restricted to their natural hosts and a few closely related species. This is thought to be due mainly to sequence divergence in the large envelope protein and species-specific differences in host components essential for virus propagation. Here we report an infection of cranes with a novel hepadnavirus, designated CHBV, that has an unexpectedly broad host range and is only distantly evolutionarily related to avihepadnaviruses of related hosts. Direct DNA sequencing of amplified CHBV DNA as well a sequencing of cloned viral genomes revealed that CHBV is most closely related to, although distinct from, Ross' goose hepatitis B virus (RGHBV) and slightly less closely related to duck hepatitis B virus (DHBV). Phylogenetically, cranes are very distant from geese and ducks and are most closely related to herons and storks. Naturally occurring hepadnaviruses in the last two species are highly divergent in sequence from RGHBV and DHBV and do not infect ducks or do so only marginally. In contrast, CHBV from crane sera and recombinant CHBV produced from LMH cells infected primary duck hepatocytes almost as efficiently as DHBV did. This is the first report of a rather broad host range of an avihepadnavirus. Our data imply either usage of similar or identical entry pathways and receptors by DHBV and CHBV, unusual host and virus adaptation mechanisms, or divergent evolution of the host genomes and cellular components required for virus propagation.


2006 ◽  
Vol 51 (2) ◽  
pp. 796-799 ◽  
Author(s):  
Ângela Novais ◽  
Rafael Cantón ◽  
Raquel Moreira ◽  
Luísa Peixe ◽  
Fernando Baquero ◽  
...  

ABSTRACT The spread of CTX-M-1-like enzymes in Spain is associated with particular plasmids of broad-host-range IncN (bla CTX-M-32, bla CTX-M-1), IncL/M (bla CTX-M-1), and IncA/C2 (bla CTX-M-3) or narrow-host-range IncFII (bla CTX-M-15). The identical genetic surroundings of bla CTX-M-32 and bla CTX-M-1 and their locations on related 40-kb IncN plasmids indicate the in vivo evolution of this element.


2018 ◽  
Vol 62 (4) ◽  
pp. e02128-17 ◽  
Author(s):  
Sead Hadziabdic ◽  
Jennie Fischer ◽  
Burkhard Malorny ◽  
Maria Borowiak ◽  
Beatriz Guerra ◽  
...  

ABSTRACT The emergence and spread of carbapenemase-producing Enterobacteriaceae (CPE) in wildlife and livestock animals pose an important safety concern for public health. With our in vivo broiler chicken infection study, we investigated the transfer and experimental microevolution of the blaNDM-1-carrying IncA/C2 plasmid (pRH-1238) introduced by avian native Salmonella enterica subsp. enterica serovar Corvallis without inducing antibiotic selection pressure. We evaluated the dependency of the time point of inoculation on donor (S. Corvallis [12-SA01738]) and plasmid-free Salmonella recipient [d-tartrate-fermenting (d-Ta+) S. Paratyphi B (13-SA01617), referred to here as S. Paratyphi B (d-Ta+)] excretion by quantifying their excretion dynamics. Using plasmid profiling by S1 nuclease-restricted pulsed-field gel electrophoresis, we gained insight into the variability of the native plasmid content among S. Corvallis reisolates as well as plasmid acquisition in S. Paratyphi B (d-Ta+) and the enterobacterial gut microflora. Whole-genome sequencing enabled us to gain an in-depth insight into the microevolution of plasmid pRH-1238 in S. Corvallis and enterobacterial recipient isolates. Our study revealed that the fecal excretion of avian native carbapenemase-producing S. Corvallis is significantly higher than that of S. Paratyphi (d-Ta+) and is not hampered by S. Paratyphi (d-Ta+). Acquisition of pRH-1238 in other Enterobacteriaceae and several events of plasmid pRH-1238 transfer to different Escherichia coli sequence types and Klebsiella pneumoniae demonstrated an interspecies broad host range. Regardless of the microevolutionary structural deletions in pRH-1238, the single carbapenem resistance marker blaNDM-1 was maintained on pRH-1238 throughout the trial. Furthermore, we showed the importance of the gut E. coli population as a vector of pRH-1238. In a potential scenario of the introduction of NDM-1-producing S. Corvallis into a broiler flock, the pRH-1238 plasmid could persist and spread to a broad host range even in the absence of antibiotic pressure.


1999 ◽  
Vol 181 (9) ◽  
pp. 2807-2815 ◽  
Author(s):  
Grazyna Jagura-Burdzy ◽  
Kalliope Kostelidou ◽  
Jessica Pole ◽  
Dheeraj Khare ◽  
Anthony Jones ◽  
...  

ABSTRACT The korAB operon of broad-host-range plasmid RK2 encodes five genes, two of which, incC andkorB, belong to the parA and parBfamilies, respectively, of genome partitioning functions. BothkorB and a third gene, korA, are responsible for coordinate regulation of operons encoding replication, transfer, and stable inheritance functions. Overexpression of incCalone caused rapid displacement of RK2. Using two different reporter systems, we show that incC modulates the action of KorB. Using promoter fusions to the reporter gene xylE, we show that incC potentiates the repression of transcription bykorB. This modulation of korB activity was only observed with incC1, which encodes the full-length IncC (364 amino acids [aa]), whereas no effect was observed withincC2, which encodes a polypeptide of 259 aa that lacks the N-terminal 105 aa. Using bacterial extracts with IncC1 and IncC2 or IncC1 purified through the use of a His6 tail and Ni-agarose chromatography, we showed that IncC1 potentiates the binding of KorB to DNA at representative KorB operators. The ability of IncC to stabilize KorB-DNA complexes suggests that these two proteins work together in the global regulation of many operons on the IncP-1 genomes, as well in plasmid partitioning.


Plasmid ◽  
1993 ◽  
Vol 29 (2) ◽  
pp. 142-146 ◽  
Author(s):  
J.M. Verger ◽  
M. Grayon ◽  
E. Chaslus-Dancla ◽  
M. Meurisse ◽  
J.P. Lafont

Plasmid ◽  
1995 ◽  
Vol 33 (1) ◽  
pp. 51-57 ◽  
Author(s):  
Philip H. Elzer ◽  
Michael E. Kovach ◽  
Robert W. Phillips ◽  
Gregory T. Robertson ◽  
Kenneth M. Peterson ◽  
...  

2022 ◽  
Author(s):  
Rahul Kaushik ◽  
Naveen Kumar ◽  
Kam Y. J. Zhang ◽  
Pratiksha Srivastava ◽  
Sandeep Bhatia ◽  
...  

Understanding the origin of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a highly debatable and unsolved challenge for the scientific communities across the world. A key to dissect the susceptibility profiles of animal species to SARS-CoV-2 is to understand how virus enters into the cells. The interaction of SARS-CoV-2 ligands (RBD on spike protein) with its host cell receptor, angiotensin-converting enzyme 2 (ACE2), is a critical determinant of host range and cross-species transmission. In this study, we developed and implemented a rigorous computational approach for predicting binding affinity between 299 ACE2 orthologs from diverse vertebrate species and the SARS-CoV-2 spike protein. The findings show that the spike protein of SARS-CoV-2 can bind to many vertebrate species carrying evolutionary divergent ACE2, implying a broad host range at the virus entry level, which may contribute to cross-species transmission and further viral evolution. Additionally, the present study facilitated the identification of genetic determinants that may differentiate susceptible from the resistant host species based on the conservation of ACE2- spike protein interacting residues in vertebrate host species known to facilitate SARS-CoV-2 infection; however, these genetic determinants warrant in vivo experimental confirmation. The molecular interactions associated with varied binding affinity of distinct ACE2 isoforms in a specific bat species were identified using protein structure analysis, implying the existence of diversified susceptibility of bat species to SARS-CoV-2. The findings from current study highlight the importance of intensive surveillance programs aimed at identifying susceptible hosts, particularly those with the potential to transmit zoonotic pathogens, in order to prevent future outbreaks.


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