Network


Latest external collaboration on country level. Dive into details by clicking on the dots.

Hotspot


Dive into the research topics where Zdenka Hradecna is active.

Publication


Featured researches published by Zdenka Hradecna.


Gene | 1996

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

Jadwiga Wild; Zdenka Hradecna; György Pósfai; Waclaw Szybalski

A prerequisite for sequencing large genomes is to obtain 30- to 150-kb genomic DNA fragments in adequate quantity. Previously, we developed a system which enables one to excise and amplify in vivo such segments directly from the Escherichia coli genome. This system, which employed the yeast Flp/FRT elements for excision and the plasmid R6K-based replication machinery for DNA amplification, permits one to bypass conventional cloning [Pósfai et al. (1994) Nucleic Acids Res. 22, 2392-2398]. To extend the applicability of such a system to many species, we describe here a broad-host-range (bhr) system in which the amplification of the excised DNA fragment depends on the oriV element and the Rep (TrfA) protein from the promiscuous RK2/RP4 plasmid. We have constructed insertion plasmids which carry the FRT and oriV sites. To introduce such plasmids into the appropriate position in the host genome, a short genomic sequence homologous to this position was cloned into the multiple cloning site (MCS) of the FRT/oriV insertion plasmid and then recombined into this position in the genome by RecA-mediated recombination. In such a manner, many strains with single FRT/oriV insertions at various positions could be generated. Subsequent genetic crosses or phage transduction allow two neighboring FRT/oriV sites (less than 150 kb apart) to be brought into a single genome. In the present report, the lacZ and phoB sites, which are 51 kb apart in the E. coli genome, were used for the introduction of the FRT/oriV sites. To deliver the Flp (excision) and Rep (amplification) functions in trans, the yeast FLP and RK2 plasmid trfA genes were placed under the control of the Ptet promoter/operator which is tightly regulated by the TetR repressor. The addition of heated chlortetracycline (cTc) inactivates TetR, turning on the synthesis of Flp and TrfA, which respectively, execute (i) excision of the 51-kb genomic segment between the two FRT sites (in lacZ and in phoB), and (ii) its amplification.


Genome Research | 2002

Conditionally Amplifiable BACs: Switching From Single-Copy to High-Copy Vectors and Genomic Clones

Jadwiga Wild; Zdenka Hradecna; Waclaw Szybalski


Virology | 1967

Fractionation of the complementary strands of coliphage T4 DNA based on the asymmetric distribution of the poly U and poly U,G binding sites.

Zdenka Hradecna; Waclaw Szybalski


Nucleic Acids Research | 1994

In vivo excision and amplification of large segments of the Escherichia coli genome

György Pósfai; Michael Koob; Zdenka Hradecna; Noaman Hasan; Marcin Filutowicz; Waclaw Szybalski


Journal of Cellular Physiology | 1969

Transcriptional controls in developing bacteriophages

Waclaw Szybalski; Kjell Bøvre; M. Fiandt; Arabinda Guha; Zdenka Hradecna; S. Kumar; Homer A. Lozeron; R. V. M. Maher; H. J. J. Nijkamp; William C. Summers; K. Taylor


Nature | 1969

Orientation and Control of Transcription in E. coli Phage λ

Sushil Kumar; Kjell Bøvre; Arabinda Guha; Zdenka Hradecna; Veronica Mary Maher; Waclaw Szybalski


Virology | 1969

Electron micrographic maps of deletions and substitutions in the genomes of transducing coliphages λdg and λbio

Zdenka Hradecna; Waclaw Szybalski


Gene | 1998

Targeting and retrofitting pre-existing libraries of transposon insertions with FRT and oriV elements for in-vivo generation of large quantities of any genomic fragment.

Jadwiga Wild; Marian Sektas; Zdenka Hradecna; Waclaw Szybalski


Archive | 2001

EXPRESSION VECTOR WITH DUAL CONTROL OF REPLICATION AND TRANSCRIPTION

Waclaw Szybalski; Jadwiga Wild; Zdenka Hradecna


Archive | 2002

Method for converting single-copy BAC vectors to conditional high-copy pBAC/oriHC vectors

Waclaw Szybalski; Jadwiga Wild; Zdenka Hradecna

Collaboration


Dive into the Zdenka Hradecna's collaboration.

Top Co-Authors

Avatar

Waclaw Szybalski

University of Wisconsin-Madison

View shared research outputs
Top Co-Authors

Avatar

Jadwiga Wild

University of Wisconsin-Madison

View shared research outputs
Top Co-Authors

Avatar

Arabinda Guha

University of Wisconsin-Madison

View shared research outputs
Top Co-Authors

Avatar

Kjell Bøvre

University of Wisconsin-Madison

View shared research outputs
Top Co-Authors

Avatar

Noaman Hasan

University of Wisconsin-Madison

View shared research outputs
Top Co-Authors

Avatar

David Frisch

University of Wisconsin-Madison

View shared research outputs
Top Co-Authors

Avatar
Top Co-Authors

Avatar

György Pósfai

Hungarian Academy of Sciences

View shared research outputs
Top Co-Authors

Avatar

H. J. J. Nijkamp

University of Wisconsin-Madison

View shared research outputs
Top Co-Authors

Avatar

Homer A. Lozeron

University of Wisconsin-Madison

View shared research outputs
Researchain Logo
Decentralizing Knowledge