The Protein Man's Blog | A Discussion of Protein Research

Loop mediated isothermal Amplification (LAMP) technique is one of the recent PCR techniques that is gaining importance for its application in molecular diagnostics. LAMP is a single tube PCR reaction carried out at constant temperature. LAMP is simple, affordable, cost effective, requires simple machinery and can be used in remote places by persons with little training.

There has been an exponential interest developing from past few years for studying the translational regulation of different genes in many model systems. Protein synthesis is the functional and fundamental biological process essential for decoding the genetic information and culminates it with a specific phenotype. Since the energy expenditure on protein synthesis is roughly more than half of the total energy expense of the cell during rapid growth, thus studying the kinetics of translational instrumentation has gained a broad attraction recently. As it has been shown previously, even a subtle altercation in translational apparatus can substantially affect human health. Hence, it is essential to profile the actively translating proteins comprehensively in order to develop insights into the cumulative translatome and its functional correlations.

Expression of recombinant proteins with peptide or protein tags is widely used in protein research for three main reasons, ease of purification from a large pool of host proteins, enhancing solubility of the protein and for localization studies. Some important steps to be considered while choosing an expression vector are compatibility of tag sequence with that of the desired protein, codon usage, including linker sequences, peptide cleavage sites and the impact of the tag on the nature of desired protein. Various tags are used ranging from large proteins (Maltose Binding Protein {MBP}) to small peptides (Hexa (6X) Histidine). Tags can be added to either N-terminal side or C-terminal side of the desired protein.

Gene cloning is one of the most important steps in recombinant DNA Technology. Now-a- days researchers are using different cloning techniques depending on purpose, time, cost, ease of use and availability of resources.  A few of the techniques are briefly explained below.