genetic typing methods of t. gondii strains have been extensively perfected in recent years. from a technical point of view, many tools usable for genetic studied on single-copy loci have been used: rflp, pcr-rflp, sequencing, rapd-pcr and isoenzyme analysis. we described the cloning and sequence analysis of the gene which encodes the major surface antigen (sag1 or p30) of t. gondii. sag1 is th...

The chloramphenicol acetyl transferase (CAT) gene is widely used as a reporter gene to analyse promoters and other gene regulatory elements. Cloning of a DNA fragment into a CAT vector such as pSVO CAT (1) usually involves blunt-end ligation with no control over the orientation of the insert. This is because the first generation CAT vectors lack convenient multiple restriction sites immediately...

A new replicon suitable for cloning and gene expression was successfully introduced into Streptococcus pneumoniae. The non-integrative lactococcal vectors pIL253 (higher-copy) and pIL252 (lower-copy), which are based on the promiscuous theta-replicating plasmid pAMβ1, were established in pneumococcus. The stability and the small size of these plasmids, together with the presence of a helpful mu...

Although controversy surrounds cloning efforts, the cloning of animals to assist efforts to preserve genetic variation in support of endangered species conservation efforts has attracted serious interest. A recent report by Loi et al. describing the cloning of a mouflon (a species of wild sheep) in a domestic sheep surrogate points to potential conservation opportunities and additional challeng...

Background & objective:  Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines is the most effective way against TB. The aim of this study was to design and construc...

BACKGROUND The selection of bacterial recombinants that harbour a desired insert, has been a key factor in molecular cloning and a series of screening procedures need to be performed for selection of clones carrying the genes of interest. The conventional cloning techniques are reported to have problems such as screening high number of colonies, generation of false positives, setting up of cont...

analyzing the gene of interest has several advantages over subcloning analysis. This method does not require any information about physical maps of the insert DNA. Positions of transposon insertion are presented as a distance from one end of the genomic DNA. Even if the restriction map is known, this technique should be preferable because of its simplicity. Using this protocol, it takes only on...

Self-cloning strains of industrial brewing yeast were constructed, in which one allele of a-acetohydroxyacid synthase (AHAS) gene (ILV2) was disrupted by integrating Saccharomyces cerevisiae genes, c-glutamylcysteine synthetase gene (GSH1) and copper resistant gene (CUP1) into the locus of ILV2. The self-cloning strains were selected for their resistance to CuSO4 and identified by PCR amplifica...

Since the mono gastric animals are not capable of metabolizing phytate, a rich source of Phosphorus, undigested phytate is excreted in manure without absorbed for metabolism and poses serious phosphorus pollution in the environment. Therefore, supplementation of phytases into animal feed is expected to solve the above problems. The present investigation deals with cloning and expression of a ph...