GPI precursor is synthesized in the endoplasmic reticulum and then is covalently attached as a posttranslational modification to the C-terminal of a protein that express a GPI attachment signal sequence . This procedure (GPI-anchor attachment) leads to stable association of the target proteins with the membrane [16, 36]. We produced a cell line that stably express ECD HER2-GPI anchored protein on cell surface. Flow cytometry and Immunoflourescent microscopy assays showed that the cell surface expression of HER2 ECD is remarkable. This cell line can frequently be used in biophysical and structural studies of binding anti HER2 agents.
The purpose of DNA extraction is to obtain DNA in a relatively purified form which can be used for further investigations such as PCR and DNA sequencing. Many different methods are available for isolating genomic DNA. Modification and optimization of DNA extraction methods are usually done for different types of cell (Dhaliwal, 2013). Cetyltrimethyl ammonium bromide (CTAB) is a preferable method that can be used for isolating the genomic DNA. There are four important steps in DNA extraction protocol of animal tissue which are enzymatic digestion of cellular protein, DNA precipitation, DNA washing, and DNA hydration (refer to Figure 1 in Appendix 1).
This was used to identify the success of EGFP ligation into pET41a(+) vector upon restriction digest and gel electrophoresis. Additionally, Polymerase Chain Reaction (PCR) was run on the isolated DNA plasmids with one of the primers specifically annealing to a part of pET41a(+) sequence and the other annealing to the EGFP gene.
The polypeptide folds and bonds with other polypeptides, and functions as a complete structure of a protein. In conclusion, DNA contains “blueprints” which are used to create cells. It provides instructions to the cell. Depending on the cells environment the genes could be switched on or to off. Since it’s the cause of the production of enzymes.
Polyacrylamide gels are usually used for proteins and for small fragments of DNA. I will focus more on the agarose gel. Agarose ... ... middle of paper ... ...gel-electrophoresis/6530 DNA Learning Center. Biology Animation Center: Gel Electrophoresis. Retrieved 12-21-2013 from Cold Spring Harbor Laboratory, website: http://www.dnalc.org/resources/animations/gelelectrophoresis.html Espionage Information.
Differential Scanning Calorimetry (DSC) The thermal properties of powdered faeces were investigated by DSC analysis because this information is important in polymer processing. The melting (Tm) and glass transition temperature (Tg) of the PHB present in the faeces was measured to be 167 °C and 3.92 °C respectively. The melting temperature of the biologically recovered PHB was found to be slightly lower when compared to values (173 - 180 °C) that were reported in the literature (Doi et al., 1995; Hahn et al., 1995; Sudesh et al., 2000).Yet, the attained value was close to the range of 160-171 °C that has been reported by Vallapil et al (2009) using different recovery methods. ... ... middle of paper ... ...g, Y. and Lee, S. (1995) Recovery and characterization of poly(3-hydroxybutyric acid) synthesized in Alcaligenes eutrophus and recombinant Escherichia coli. Appl Environ Microbiol 61, 34-39.
What is important when you design primers? there are several considerations for primers design including primer length between 18 and 24 bases, Melting Temperature (Tm) , 3' sequence and guanine-cytosine content .  3. Describe how PCR is performed! ● step 1- DNA , tow primer (forward and reverse primer), dNTPs , and DNA polymerase enzyme mixed together in sutable tube.
Protein Ontology (PRO) is a curation that was designed as a Open Biomedical Ontologies (OBO) Discoveredry ontology for proteins. It contains information on the classification of proteins, evolutionary relationships and the multiple protein forms of a gene. These components include post-translational modifications, alternative splicing and cleavage. This paper focused on the TGF-beta signaling proteins and specifically described the usage, curation, building and dissemination of PRO. PrePRO serves as the precursor to PRO, which was manually curated.
DNA methylation in such genomes can affect cytosine residues in 3 different contexts: CG, CHG and CHH, where H can be C, T or A. One can wonder how these methylations are maintained in plant genomes. This can be explained by the dimethylation activity of cytosine-DNA methyltransferases on histone H3 at lysine 9(H3K9). DNA methylation is initiated by RNA-directed methylation (RdDM), a process which is led by small and long non-coding RNAs via the Dicer-Argonaute pathway (Dinh et al., 2013). Once DNA methylation is initiated, it must be maintained to effectively suppress gene transcription in heterochromatin.
On the other hand RNA (ribonucleic acid) contains nucleotides having sugar ribose and is usually single stranded. RNA contains the bases guanine, cytosine, adenine, and uracil. Some of the components of RNA that allows it to function are have the components of ribosomes (rRNA), transporting amino acids (tRNA), and translating the message of the DNA code (mRNA) (Freeman). The first step in converting or copying genetic information is by the process called transcription. Transcription occurs in the nucleus of the eukaryotic cell and begins when the sigma is binded to the promoter of the start of the gene (Freeman).