Aqueous two-phase system (ATPS) is a liquid-liquid fractionation technique and has gained an interest because of great potential for the extraction, separation, purification and enrichment of proteins, membranes, viruses, enzymes, nucleic acids and other biomolecules both in industry and academia. Although, the partition behavior involved in the method is complex and difficult to predict. Current research shows that it has also been successfully used in the detection of veterinary drug residues in food, separation of precious metals, sewage treatment and a variety of other purposes. The ATPS is able to give high recovery yield and is easily to scale up. It is also very economic and environment friendly method. The aim of this review is to overview the basics of ATPS, optimization and its applications.
The development of antibiotic resistance in bacteria is a major public health threat. Infection rates of resistant pathogens continue to rise against nearly all antimicrobials, which has led to development of different strategies to combat the antimicrobial resistance. In this review, we discuss how the newly popular CRISPR-cas system has been applied to combat antibiotic resistance in both extracellular and intracellular pathogens. We also review a recently developed method in which nano-size CRISPR complex was used without any phage to target the
mecA
gene. However, there is still challenge to practice these methods in field against emerging antimicrobial resistant pathogens.
Food contamination with antibiotic residues has become a worldwide problem due to the heavy and improper use of antibiotics. The development of fast, high-throughput, easy, and economical screening methods for the detection of antibiotic residues is an important requirement as an alternative to the traditional assays. Immunoassays are widespread promising methods that can achieve the conditions of an analytical method in the evaluation of food and environmental protection. This review offers an overview of the preparation of antibodies and the currently established immuneassays for the antibiotic residues detection, including enzymelinked immunosorbent assay, fluorescence immunoassay, radioimmunoassay, colloidal gold immunoassay, and chemiluminescence immunoassay. Furthermore, it will elaborate on the future perspective on the performance and improvement of these assays for the determination of antibiotic residues in food samples. These assays include a simple sample preparation and high-sensitive detection that offer a significant assurance for the screening of antibiotic residues.
Prokaryotes have developed numerous innate immune mechanisms in order to fend off bacteriophage or plasmid attack. One of these immune systems is clustered regularly interspaced short palindromic repeats (CRISPR). CRISPR-associated proteins play a key role in survival of prokaryotes against invaders, as these systems cleave DNA of foreign genetic elements. Beyond providing immunity, these systems have significant impact in altering the bacterial physiology in term of its virulence and pathogenicity, as well as evolution. Also, due to their diverse nature of functionality, cas9 endoribonuclease can be easily reprogrammed with the help of guide RNAs, showing unprecedented potential and significance for gene editing in treating genetic diseases. Here, we also discuss the use of NgAgo–gDNA system in genome editing of human cells.
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