Lvb Lab Report

The vital components and techniques of gene cloning are as follows, the DNA sequence that contains the desired gene (EZH2) is amplified by Polymerase chain reaction. PCR was established by Kary Mullis in 1985, popularly known to amplify target sequences of DNA (EZH2) to a billion fold in several hours using thermophilic polymerases (Taq) ,primers and other cofactors (Sambrook and Russell, 2001). Three crucial steps are involved which are Denaturation (at 95°), Annealing of the forward and reverse primers (55-65°) and lastly primer extension (at 72°). After amplification the desired sequence is integrated into the circular vector (pbluescript) forming the recombinant molecule. For the compatibility of the insert and vector, both were digested with (EcoR1) so the same cohesive ends are generated in both, making it easier to ligate. EcoR1 is a restriction enzyme that belongs to the type II endonuclease class which cuts within dsDNA at its recognition site “GAATTC” (Clark 2010; Sambrook and Russell, 2001).

To construct a mapping cross of linked genes, it is important that the genotypes of all of the

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 polymeric chains that are built from monomers called amino acids. All structural and functional properties of proteins derive from the chemical properties of the polypeptide chain. There are four levels of protein structural organization: primary, secondary, tertiary, and quaternary. Primary structure is defined as the linear sequence of amino acids in a polypeptide chain. The secondary structure refers to certain regular geometric figures of the chain. Tertiary structure results from long-range contacts within the chain. The quaternary structure is the organization of protein subunits, or two or more independent polypeptide chains.

Snustad, D. P. and Simmons, M. J. Principles of genetics 6th Ed. (John Wiley & Sons, Inc., New York, 2012).

The active site of the protein is located at the cleft between the subdomains. It is characterized as a mix of type 1 and type 2 RIPs. Around the N-terminus, there are 6 β-sheets, 4 antiparallel strands and 2 parallel strands. The C-terminus has 8 α-helices where the 310-helix crosses over on the parallel β-strand giving a canonical geometry. Unlike RIPs, Saporin-S6 has 2 β-strands (7 & 8) that are shorter that increases the accessibility to a particular substrate. This plays a role with its resistance to denaturing agents, the ability to survive in extreme pH, the modifications that are necessary to create immunotoxin and the stability in extreme temperatures.

The F2 generation of this test group will then be screened for the roller mutation. The progeny of these rollers, the F3 generation, will yield the most integral data of our screen.

One of the central themes of biology is that structure and function are tied together.Typically, when a researcher looks at an entity such as a biomolecule, one of the most useful approaches is to gather information about its structure in order to gain insight to its functions and properties.

With an algorithm we can locate the motif much faster to analyze it. Once the motif is found,

RFLP or restriction fragment length polymorphism method is used in which restriction enzyme cuts the DNA at specific sites according to different restriction enzymes. Length of fragments will be different for each individual (everyone’s DNA is unique).

The motif was compared with its ancestral sequence to test, and further validate, whether it is a good binding motif. The similarities between and ancestral were compared to the ones between motif and random promoter sequences. The results showed that in most cases, a good biding motif was embedded in an ancestral repeat instead of a random promoter sequence; this confirms that the ancestral repeats were more easily able to produce a good binding

Punnett squares are grid-like charts that is used to predict the probability of genotype of offspring. Biologists use this chart to see the probability of an offspring having a particular genotype. The chart consists genotypes, which are genes, and phenotype,which are physical. There’s also a two factor crosses which track two traits at a time. To set up the gametes in a two factor cross you need to list all the possible combinations of the alleles by using FOILing.

We have the sequences and the PDB files for all proteins that have been successfully aligned with active peptides. We design self-inhibitory peptides targeting these proteins using SIP. The process consists in four steps that will be briefly described here: The first step is the secondary structure prediction followed by the second step, which is the disorder prediction. These first two steps are based on the protein sequence only. The third step is the selection of segments of 20 amino acid long, with low disorder and helical secondary structure, within the protein. And finally, the fourth step is the calculation of the the energy score based of the structural information from the PDB file.

The structure of the prepared material was investigated employing many techniques such as X-rays diffraction, SEM and TEM electron microscopy. In order to scrutinize the crystal structure of the samples X-rays diffraction was carried out using diffractometer XPERT-MPDUG, Philips PW3040. Figure (1) depicts the X-ray diffraction of the as synthesized samples. It can be seen that the nature of the sample is formed in poly crystalline form. The most important peaks are displayed. The most intense peaks were indexed to be 201, 011, 111, 400, 311, 102, 020, and 112. The indexed peaks correspond to single pure orthorhombic phase according to ICDD card 83-1758 (75-2115). The reflection planes, regarding the ICDD card, are

To investigate the efficacy of proposed feature extraction approach, the experimentation is performed on two superfamilies, namely Ras and Globin. Across tenfold cross validation, the highest Classification Accuracy and Computational Time achieved by proposed approach is 94.323.52 and 6.54(s) respectively in comparison to the Classification Accuracies achieved by other approaches [4], [5] and [6] are 85.420.55, 67.518.38, 51.410.27 with Computational Time 7.11(s), 10.13(s), 63.98(s) respectively. The experimental results demonstrate that the proposed approach extract the minimum relevant features for each protein sequence.