BACKGROUND In vivo recombination of overlapping DNA fragments for assembly of large DNA constructs in the yeast Saccharomyces cerevisiae holds great potential for pathway engineering on a small laboratory scale as well as for automated high-throughput strain construction. However, the current in vivo assembly methods are not consistent with respect to yields of correctly assembled constructs an...

Transient gene expression (TGE) in mammalian cells is a method of rapidly generating recombinant protein material for initial characterisation studies that does not require time-consuming processes associated with stable cell line construction. High TGE yields are heavily dependent on efficient delivery plasmid DNA across both the plasma and nuclear membranes. Here, we harness nucleoside diphos...

BACKGROUND RSF1010 is a well-studied broad-host-range plasmid able to be mobilized to different bacteria and plants. RSF1010-derived plasmid vectors are widely used in both basic research and industrial applications. In the latter case, exploiting of mobilizable plasmids or even the plasmids possessing negligible mobilization frequency, but containing DNA fragments that could promote conjugal t...

The recent Zika virus (ZIKV) outbreak has been linked to severe pathogenesis. Here, we report the construction of a plasmid carrying a cytomegalovirus (CMV) promoter-expressed prototype 1947 Uganda MR766 ZIKV cDNA that can initiate infection following direct plasmid DNA transfection of mammalian cells. Incorporation of a synthetic intron in the nonstructural protein 1 (NS1) region of the ZIKV p...

A region of DNA which determined the production of the insecticidal toxin of Bacillus thuringiensis subsp. israelensis was cloned into a derivative of a broad-host-range group IncQ plasmid vector of gram-negative bacteria. The plasmid which we constructed was transferred by conjugative mobilization into a Bradyrhizobium species that nodulates pigeon peas. In this species the construction was ma...

Methods pPRV111A::neoSTLS2 construction and transplastomic plants pPRV111A::neoSTLS2 was produced by ligating the HindIII fragment containing neoSTLS2 from plasmid pCMneoSTLS2 (ref. 13) into pPRV111A. This transformation vector was introduced into the plastids of Nicotiana tabacum L. Petit Havana (N,N) by biolistic bombardment, and homoplasmic plants were regenerated as described previously. Th...

conclusions based on our findings, p. aerugionsa is a major causative agent of wound infections and amikacin could be considered as a more effective antibiotic for treatment of the burned patients. background nowadays, the treatment of burned patients is difficult because of the high frequency of infection with antibiotic resistance bacteria. objectives this study was conducted to evaluate the ...

conclusions: the recombinant baculovirus constructed in this work has proper characteristics to produce h5n1 influenza virus-like particles in sf9 cells. results: restriction map of pfastbacihnm1 plasmid confirmed the fidelity of the clone. the pcr carried out on the recombinant bacmids as template indicated that a proper homologous recombination has occurred between pfastbacihnm1 donor plasmid...

Derivatives of the Pseudomonas aeruginosa plasmid R91-5, loaded with the transposon Tn501, were transferred to P. putida PPN. Over 90% of exconjugants, which arose at a frequency of ca. 10(-6) per donor cell, exhibited high-frequency (greater than 10(-2) per donor cell) polarized transfer of chromosomal markers. In one instance it was demonstrated by transduction that the plasmid had been inser...