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Vaccines, Hepatitis B and Insulin Production
Published in Debabrata Das, Soumya Pandit, Industrial Biotechnology, 2021
Our body relies on protein production by using transcription and later translation, but in vitro this can be achieved by Recombinant DNA Technology (RDT), which will produce recombinant proteins (Brown et al., 1993). Plasmids are small circular DNA molecules which are present on bacteria which can transmit to daughter cells by methods such as cell division, binary fission or conjugation. Scientists have devised a novel method which uses Restriction Enzymes (RE) to cleave the insert as well as the plasmid, which is used to introduce the protein-coding gene into the plasmid, growing them in culture to maximize the amount of desirable genes (Figure 13.2). The target genes are then introduced into host cells or vectors which possess specialized machinery for transcription and translation, which represents an expression system.
Impact of Massively Parallel Computation on Protein Structure Determination
Published in Theo C. Pilkington, Bruce Loftis, Joe F. Thompson, Savio L-Y. Woo, Thomas C. Palmer, Thomas F. Budinger, High-Performance Computing in Biomedical Research, 2020
Richard C. Brower, Charles DeLisi
The challenge we describe here is not just of fundamental biological importance; it is central to the success of the biotechnology industry and hence to national competitiveness. Proteins produced by recombinant DNA technology often do not fold properly. In the absence of improved understanding of protein folding, progress on effective protein production is likely to be slow. In addition, a predictive understanding of protein and peptide structure is of fundamental importance to rational approaches to the design of drugs and vaccines. The challenge thus links two of America’s most promising technologies: biotechnology and computer engineering.
Protein Expression Methods
Published in Jay L. Nadeau, Introduction to Experimental Biophysics, 2017
A wide variety of different expression strains and cell lines exist for protein production, with different strains or lines providing application-specific advantages. Here, the common expression strains and cell lines used for each of the various expression systems outlined in Section 5.2, “Expression Systems” section are briefly reviewed, focusing on bacterial expression systems.
Polyclonal antibody production against rGPC3 and their application in diagnosis of hepatocellular carcinoma
Published in Preparative Biochemistry and Biotechnology, 2018
Shenghao Wang, Muhammad Kalim, Keying Liang, Jinbiao Zhan
Escherichia coli expression system possesses a potent source of recombinant protein production. But in the expression of exogenous protein, especially in the expression of eukaryotic proteins, there are still many problems that limit expression level, no expression or lost biological activity due to structural change and protein modification translation. We have achieved the high expression level of GPC3 N-terminal protein by deleting the 18 bases of six proline residues from GPC3 N-terminal protein. Successive washing methodology and refolding techniques provide an active production of recombinant proteins. Thereafter, the rGPC3 N-terminal protein was used as an antigen to immunize New Zealand male rabbits to obtain anti-GPC3 polyclonal antibody. The polyclonal antibody has shown specific binding interactions to the GPC3 surface antigen on primary liver cancer cell lines (Huh7 and HepG2). Immunohistochemistry assay also proves that polyclonal antibody can combine with GPC3 in HCC cancer tissue. In addition, the recombinant GPC3 N-terminal protein provides a basis for the screening of monoclonal antibodies for HCC-targeted therapy. The findings here will be useful for the production of valuable antibodies which can be used for cancer diagnosis and immunotherapy.
Application of Meta-Heuristics on Reconstructing Gene Regulatory Network: A Bayesian Model Approach
Published in IETE Journal of Research, 2021
Suman Mitra, Surama Biswas, Sriyankar Acharyya
Each gene codes a protein. The process of protein production is named as gene expression. The degree of gene expression at a particular time is called gene expression level [1]. Protein produced from one gene influences other gene(s) to change their protein production rate. As a result of gene–gene interaction, a network is constituted where genes are the nodes and the influence of a gene to other is an edge. This network is termed as Gene Regulatory Network (GRN) [2]. Reconstruction of GRN is very important to analyse the behaviour of the normal and diseased cells.
Cloning, expression and characterization of a HER2-alpha luffin fusion protein in Escherichia coli
Published in Preparative Biochemistry and Biotechnology, 2019
Farzaneh Barkhordari, Nooshin Sohrabi, Fatemeh Davami, Fereidoun Mahboudi, Yeganeh Talebkhan Garoosi
Protein expression is a multifactorial process in which the amount of expressed protein per cell and cellular mass per volume of the medium will be changed. Therefore, optimization of conditions is a crucial step in large scale protein production.[35] Several factors influencing cellular growth and recombinant protein expression such as medium type, incubation temperature, inducer concentrations and time of post-induction were evaluated in this study.