the clar of streptomyces clavuligerus in the clavulanic acid gene cluster encodes a transcriptional regulator that controls clavulanic acid biosynthesis. the main goal of this study was isolation and molecular detection of the clar gene and its cloning in the streptomyces specific vector (pma:: hyg). by cinsideration of the clar gene’s start codon, the specific primers were designed. after geno...

In vivo recombinational cloning in yeast is a very efficient method. Until now, this method has been limited to experiments with yeast vectors because most animal, insect, and bacterial vectors lack yeast replication origins. We developed a new system to apply yeast-based in vivo cloning to vectors lacking yeast replication origins. Many cloning vectors are derived from the plasmid pBR322 and h...

An efficient method for constructing a recombinant adenovirus (Ad) vector, based on an in vitro ligation, has been developed. To insert the foreign gene into an adenoviral DNA, we introduced three unique restriction sites, I-CeuI, SwaI, and PI-SceI, into the E1 deletion site of the vector plasmid, which contains a complete E1, E3-deleted adenovirus type 5 genome. I-CeuI and PI-SceI are intron-e...

We have developed a method to improve the transformation efficiency in genome-sequenced bacteria, using 'Plasmid Artificial Modification' (PAM), using the host's own restriction system. In this method, a shuttle vector was pre-methylated in Escherichia coli cells, which carry all the putative genes encoding the DNA modification enzymes of the target microorganism, before electroporation was per...

In the present study, the bovine coronavirus (BCV) spike glycoprotein cleavage products (S1 and S2) were individually expressed in Spodoptera frugiperda (Sf9) insect cells, using a baculovirus expression system. The coding sequence of S1 and S2 gene fragments were amplified by RT-PCR and cloned into the baculovirus shuttle vector pBlueBac4.5/V5-His TOPO ® TA. The cloned fragments were inserted ...

collagens are the most abundant proteins in the human body. their main function is to provide structural and mechanical support for the tissues, but they are also involved in a number of other biological functions including cell attachment, migration and differentiation. collagens and gelatins are widely used in pharmaceutical and medical applications. every year, more than 50,000 tons of colla...

The cyanobacteria are an ancient group of Gram-negative prokaryotes. In addition to photosynthesis, the cyanobacteria possess many other interesting features: a) nitrogen-fixation, b) cellular differentiation, c) regulated genomic rearrangements (1, 2) and, d) introns (3). Studies of these features in the filamentous cyanobacteria are hampered by a lack of shuttle vectors. Currently, there is o...

background: leishmania major lmsti1 is a conserved protein among different species of leishmania, and expressed in both amastigote and promastigote forms of l. major life cycle. it has previously been expressed in bacterial systems. m aterials and methods: to express lmsti1 in the methylotrophic yeast         pichia pastoris ( p. pastoris ), the shuttle vector ppicza containing gene lmsti1 was ...