Restriction enzymes represent a class of endonucleases that cleave DNA in a specific manner. They are widely used in recombinant DNA technology and known as molecular scissors. In this communication, a putative restriction endonucleases from (Acinetobacter baumannii SDF) was identified in NCBI database, and characterized for biophysical properties, sequence homology and a structural modeling. T...

Most restriction endonucleases use Mg2+ to hydrolyze phosphodiester bonds at specific DNA sites. We show here that BfiI, a metal-independent restriction enzyme from the phospholipase D superfamily, catalyzes both DNA hydrolysis and transesterification reactions at its recognition site. In the presence of alcohols such as ethanol or glycerol, it attaches the alcohol covalently to the 5' terminus...

Three approaches were used to identify DNA polymorphisms in humans by examining the DNAs of a number of individuals within Huntington's Disease pedigrees as well as random individuals for restriction fragment length polymorphisms (RFLPs). The first approach used randomly chosen, cloned single-copy DNA segments from four types of human recombinant DNA libraries as hybridization probes to detect ...

Partial digestion of DNA fragments is a standard procedure for subcloning analysis and for generating restriction maps. We have developed a novel method to generate a partial digestion for any DNA fragment that can be amplified by PCR. The method involves the incorporation of 5-methyl-dCTP into the PCR product to protect most of the restriction sites. As a result, complete digestion of the modi...

Precise DNA manipulation is critical for molecular biotechnology. Restriction enzyme-based approaches are limited by their requirement of specific enzyme sites. Restriction-free cloning has greatly improved the flexibility and speed of precise DNA assembly. Most of these approaches focus on DNA assembly rather than gene removal. Here we present a polymerase chain reaction (PCR)-based cloning me...

toxoplasmosis is an infection caused by the protozoan parasite toxoplasma gondii (t.gondii) throughout the world. although usually asymptomatic, the infection can cause serious medical problems in immunocompromised individuals and fetuses. toxoplasmosis also causes considerable economic loss because of abortion in livestock. dna vaccination is a promising approach against intracellular parasite...