Antibiotic resistance chimeric plasmids have been constructed by in vitro enzymatic manipulation and introduced into Bacillus subtilis by transformation. The parental plasmids used had been introduced into B. subtilis from Staphylococcus aureus by transformation. Of the seven recombinant plasmids constructed using restriction endonucleases, one was made using EcoRI, another using Hpa II, and fi...

Phi-3-T is a Bacillus subtifis temperate bacteriophage, isolated by Tucker (11) from the soil. Tucker has shown that infection by this phage causes, by lysogenic conversion of thyB. subtifis clones, prototrophy. The phage carries genetic information specifying the thymidylate synthetase activity, the thyP gene. Recently, we have reported purification of B. subtilis DNA segments achieved by EcoR...

The replication region of the Bacillus thuringiensis plasmid, pHT1030, was treated with hydroxylamine. Various copy-number mutants were selected and subsequently used to construct shuttle vectors with multiple cloning sites. These recombinant plasmids are very stable and allowed the cloning of a delta-endotoxin-encoding gene in B. thuringiensis. Comparison between gene expression level and vect...

Interferon is a protein secreted by eucaryotic cells following stimulation by viruses, bacteria, and many other immunogenes. Recent medical studies indicate that interferons have effective role in the treatment of virus infections, immunodeficiency and certain types of cancer such as hairy cell leukaemia (HCL). The aim of the present study is to apply yeast strain for secreting human IFNα2b fol...

background: mycobacterium tuberculosis is the causative agent of tuberculosis (tb). bacille calmette-guerin (bcg) vaccine, is not effective in adults, therefore, many efforts have been made to produce an effective adult tb vaccine. the aim of this study was to develop a new tuberculosis dna vaccine candidate encoding a recombinant hspx-ppe44-esxv fusion antigen of m. tuberculosis. methods: a fu...

The use of HPLC in the separation of DNA fragments is well established (1). We have adapted these techniques to the separation of cDNA from linker sequences present during lambda cloning. Short linker sequences containing an EcoRl restriction site are coupled to blunt ended cDNA, excess linkers are cleaved using EcoRl. The efficiency of cloning into the lambda EcoRl site is improved by the remo...

Genetic studies with Burkholderia cepacia complex isolates are hampered by the limited availability of cloning vectors and by the inherent resistance of these isolates to the most common antibiotics used for genetic selection. Also, some of the promoters widely employed for gene expression in Escherichia coli are inefficient in B. cepacia. In this study, we have utilized the backbone of the vec...

PCR fragments and linear vectors containing overlapping ends are easily assembled into a propagative plasmid by homologous recombination in Escherichia coli. Although this gap-repair cloning approach is straightforward, its existence is virtually unknown to most molecular biologists. To popularize this method, we tested critical parameters influencing the efficiency of PCR fragments cloning int...

The use of gene fusion technologies for transcriptional analysis of gene expression, especially of those genes whose products are difficult to assay, has been well documented (12–15). Many narrow and broad-host-range plasmid-based systems are available for constructing such gene fusions (6,7, 14). To circumvent some of the inherent problems of plasmid-based systems, which have been discussed be...