Protein Synthesis Essay

Then the tRNA molecules link together and transfer the amino acid to the ribosome. An Anticodons pair with a codon takes the

VOCABULARY: Without using the internet, write the definition for the words you know. Then if you have blanks look them up in your textbook. (middle of the book A)

With two T7 promoters, the PCR product will be transcribed by the T7 polymerase. Therefore, a double stranded RNA (dsRNA) product is produced from the plasmid, once IPTG is added to activate the lac operator of T7 polymerase.

A protein has multiple existing structures, these are the primary, secondary, tertiary and quaternary structures which occur progressively. A protein is essentially a sequence of amino acids which are bonded adjacently, and interact with one another in various ways depending on the R group that the amino acid contains. There are 20 different amino acids which are able to be arranged in any given order, thus giving rise to a potential 2.433x1018 (4.s.f) different combinations, and therefore interactions between the various amino acids.

Proteins are primarily considered to have one primary function to serve its role in an organism, however studies have observed to have multiple functioning proteins known as moonlighting proteins (Khan et al. 2014). Moonlighting proteins along with primary functions, have secondary functions that are not related to the primary function and does not correlate to the primary or other functions (Khan et al. 2014). The multifunctional proteins play essential roles in carrying out biochemical functions which aids in the cell growth but are not caused by gene fusion and multiple RNA splice variants (Amblee et al. 2015). The discovery of moonlighting proteins was first discovered by Piatigorsky and Wistow while observing crystallins (Khan et al. 2014). Crystallins, are structural proteins that are found in the eye lens that exhibit enzymatic activity to make the lens itself (Khan et al. 2014). Crystallin has a primary function to help form the lens of the eye by acting as a structural protein (Amblee et al. 2015). Besides enzymatic activity, crystallin was observed in other mammals to have secondary functions such as metabolic functions which are helpful in prokaryotic (Khan et al 2014). Most moonlighting proteins are characterized as cytosolic enzymes and chaperons, or in other words helping proteins (Amblee et al 2015). The multifunctional proteins or moonlighting proteins can also be identified as receptors, channel proteins and ribosomal proteins (Khan et al. 2014). Due to the

These proteins differ depending on their function in the cell, but often prevent misfolding of nascent protein, and assist in refolding of the misfolded proteins. Protein folding is dependent on the amino acid sequence within the polypeptide chain that is synthesized from the DNA strand in the ribosome.2 Upon release from the ribosome, the polypeptide chain undergoes a series of conformational changes based on the amino acid sequence to produce a functional protein structure. The assembled native protein is directed to the ER via vesicle

This is done by means of the aminoacyl attachment site (the site at which the amino acid is attached to the tRNA molecule). Each tRNA molecule, by means of their anticodons (a sequence of three exposed free bases complimentary to that of the codons on

The Functions of Proteins Introduction Protein accounts for about three-fourths of the dry matter in human tissues other than fat and bone. It is a major structural component of hair, skin, nails, connective tissues, and body organs. It is required for practically every essential function in the body. Proteins are made from the following elements; carbon, hydrogen, oxygen, nitrogen and often sulphur and phosphorus.

A simple but effective method is correctly designing gRNAs. Ideally, the target sequence must be a unique and nearly perfect homology to the gRNA with as little to no homology at other points within the gene sequence. Another potential strategy involves reducing the concentration of gRNA and Cas9 that are expressed in cells. However, this strategy is considered inconclusive due to inconsistent reporting. Some cell lines showed a large decrease in rates of off-target binding while others showed no effect. An increasingly promising method for improving specificity involves using a mutated nickase version of the Cas9 enzyme. This modified version contains only one catatylic domain and is used as a single nickase system capable of cutting a single strand of DNA. This results in a “nick” or single stand break rather then a DSB and is repaired by HDR. To further reduce the risk of off target binding, a double or dual nickase CRISPR system can be implemented. This system uses two nickases which bind to opposite strands of the target gene, generating a DSB. This system closely resembles the Fokl enzyme activation used in TALEN, in that off target effects are reduced because the probability of two off target nicks within the same region is very unlikely. When the DSB is made, activation of NHEJ is initiated; to further increase specificity and reduce off-target effects, the HDR mediated editing is advised for both nickase

Genes are the foundation of life. Biologists throughout history have studied genes with the goal of knowing how they work, what functions specific genes have, and what they can do with this knowledge to further science and society. Techniques involving genes have grown to the level where we can target specific genes to edit them and even alter their function. Some earlier techniques of this include meganucleases, zinc finger nucleases, and transcription activator-like effector nucleases, but each of these techniques has its disadvantages (Sander and Joung, 2014). Recently in the world of genetics, a new and exciting method to target specific genes easily and with high precision has emerged. Clustered regularly interspaced short palindromic

Enzymes are an important part of all metabolic reactions in the body. They are catalytic proteins, able to increase the rate of a reaction, without being consumed in the process of doing so (Campbell 96). This allows the enzyme to be used again in another reaction. Enzymes speed up reactions by lowering the activation energy, the energy needed to break the chemical bonds between reactants allowing them to combine with other substances and form products (Campbell 100). In this experiment the enzyme used was acid phosphates (ACP), and the substrate was p-nitrophenyl phosphate.

Tryptophan is an amino acid used for the synthesis of proteins. An example of a bacterium that takes up the tryptophan and uses it to build proteins is Escherichia coli. This bacterium can synthesize tryptophan using enzymes that are responsible for producing five genes that are located next to each other in a tryptophan (trp) operon. An operon consists of operator, promoter and the five genes it controls. The trp operon from E.coli is normally is “on” and the genes for tryptophan synthesis are transcribed. The E.coli only uses enzymes to synthesize tryptophan when there is none in the medium but if it is present, then the enzymes are no longer needed and the operon is turned off by a protein repressor. The protein repressor does this by

In most instances, protein molecules are usually embedded from hundreds to thousands of amino acids. A repertoire of twenty different amino acids, joined in any possible sequence allows the existence of an inconceivably large number of proteins that is infinite in nature.

Protein synthesis is one of the most fundamental biological processes. To start off, a protein is made in a ribosome. There are many cellular mechanisms involved with protein synthesis. Before the process of protein synthesis can be described, a person must know what proteins are made out of. There are four basic levels of protein organization. The first is primary structure, followed by secondary structure, then tertiary structure, and the last level is quaternary structure. Once someone understands the makeup of a protein, they can then begin to learn how elements can combine and go from genes to protein. There are two main processes that occur during protein synthesis, or peptide formation. One is transcription and

RiboTALE networks studied were comprised of two modules and each module was based on a separate plasmid. One of the modules, repressor module, expresses fusion of riboswitch (RS) from an inducible PBAD promoter, second module, target module, contains a TALE binding site (TBS) downstream of a regulated or constitutively expressed promoter. Ribosome binding site (RBS) and gfp are present downstream to TBS (Fig. 1). gfp was used as proxy to quantitate the promoter expression. Repressor modules were based on kanamycin resistance medium copy number BioBrick plasmid, pSB3K3 (Shetty et al, 2008). Four different types of repressor modules, PR1T1, PR1T8, PR2T1, and PR2T8, were constructed combinatorially by implementing two types of riboswitches and two types of TALEs (supplementary table 1). Used riboswitches, Riboswitch-1 and 2, were theophylline inducible (Lynch & Gallivan, 2009; Topp et al, 2010). TALES with either low dissociation constant (Kd = 1.3±0.3, T8) or with high dissociation constant (Kd= 240±40, T1) were selected (Meckler et al, 2013). Each of the TALEs recognizes a specific 19 bp target DNA sequence.Two families of target modules were constructed. One of the modules was based on regulated tetracycline promoter, PTet, and other was based on a family of constitutive promoter, PConst, generating different level of outputs. Target modules contain two types of TALE binding site, TBS1 or TBS8. TALE1 binds with TBS1 and TALE8 binds with TBS8 (Fig.1). All measurements were