Cloning, Expression, Characterization and Purification of Endoglucanase Gene

Cloning, Expression, Characterization and Purification of Endoglucanase Gene

Length: 2561 words (7.3 double-spaced pages)

Rating: Research Papers

Open Document

Essay Preview

DISCUSSION
Endo-1, 4-B-glucanase is an important enzyme from industrial point of view and its production is considered essential for successful utilization of cellulosic materials (Wu and Lee 1997., Zhang and Lynd 2004). The objective of present research work was cloning, expression, purification and characterization of endoglucanase (bgl C) gene obtained from strain of Bacillus licheniformis ATCC 14580. Isolation of high quality genomic DNA is a prerequisite for molecular techniques (Ansari et al., 2012). The reliability of the method used for the isolation of DNA from biological samples or from reaction mixtures is of significant importance. In the present study a simple DNA isolation protocol developed by Kronstad et al (1983) was used because of its simplicity and cost effectiveness. This method was fast and complete DNA extraction was achieved within 2 h. Many workers have used other methods for the isolation of genomic DNA (Errington, 1984, Ausubel et al., 1987 and Hong et al., 2005). Amplification of endoglucanase gene was performed by polymerase chain reaction using specific primers (forward and reverse) each complementary to one end of target sequence. Primers were designed using software vector NTI. In forward primer, NdeI site a type II restriction endonuclease purified from Neisseria denitrificans was added. NdeI, is a four base cutter with recognition site at 5’-CA↓TATG-3’ and creates overhangs in cut DNA fragments (Chang et al., 2005). It creates “ATG” at the beginning of the gene from where the transcription will start in the cloned gene. Ganiger et al., 2008 also amplified β-1, 6 endoglucanase gene from T. harzianum using PCR. Meera et al., 2011 amplified an intronless gene encoding endoglucanase eng 61 from Aspe...


... middle of paper ...


...arameters and optimized culture conditions which maximize enzyme yield. As far as the expression level of endoglucanase gene is concerned our results are much better than the recently conducted studies on the expression of endoglucanase of various organisms (Shyamala et al., 2011; Akita et al., 2005; Noronha and Ulhoa 2000). Endoglucanase is an industrially important enzyme involved synergistically in degradation of cellulosic materials into constitutive monomers with other cellulase enzymes. Its broad pH range and higher temperature tolerant ability make it useful for various industrial processes that are proceeded at higher temperature as well as can also help in cutting down cost of various processes. Because of the advantageous properties demonstrated by this enzyme, it can be further exploited for enzymatic hydrolysis of locally produced agriculture biomass.

Need Writing Help?

Get feedback on grammar, clarity, concision and logic instantly.

Check your paper »

Comparative Studies Of Male And Female Gene Expression Essay

- Recent comparative studies of male and female gene expression profiles (Kobayashi et al. 2006; Gutiérrez-Adán et al. 2006) called into question the validity of the dominating idea assuming that phenotypic sex differences in higher vertebrates appear only with the start of sex hormone secretion by the gonads (Jost 1947). Faster developmental rate of the pre-implantation male embryos and higher viability of the female embryos are the most noticeable differences between the sexes (Gutiérrez-Adán et al....   [tags: Sex, Male, Gene expression, Gender]

Research Papers
1003 words (2.9 pages)

Essay about A Research Study On Gene Therapy

- Researchers are becoming more proficient at using genes to help change the effects of some diseases, potentially opening the door to curing diseases that have ravaged humankind for centuries. Holden Gowan is an undergraduate student at the University of North Carolina at Chapel Hill with a plan to major in chemistry with a focus in biochemistry Introduction Imagine living in a world where diseases like hemophilia, cancer, cystic fibrosis, and AIDS did not exist. This particular world might not be as far away as one might think due to a relatively new type of treatment known as gene therapy....   [tags: DNA, Gene, Gene expression, Genetics]

Research Papers
939 words (2.7 pages)

A Research Study On Gene Therapy Essay

- The first video in the assignment was about three people who suffer from a genetic mistake that has led to their diseases. The video also gives background on the human genome. The human genome was sequenced about ten years before the film was made. The three people in this video are Sophie, Emma, and Tom. Sophie suffers from cystic fibrosis, Emma suffers from breast cancer, and Tom suffers from alcoholism. These three people went on a journey to learn new facts about their disease and treatment plans....   [tags: DNA, Gene, Genetics, Gene expression]

Research Papers
954 words (2.7 pages)

Being Treated With Gene Therapy Essay

- Holden Gowan is an undergraduate student at the University of North Carolina at Chapel Hill with a plan to major in chemistry with a focus in biochemistry Introduction Imagine living in a world where diseases like hemophilia, cancer, cystic fibrosis, and AIDS did not exist. This particular world might not be as far away as one might think due to a relatively new type of treatment known as gene therapy. All of the aforementioned diseases deal with problems concerning genes. Gene therapy is able to correct the problems that have forced one to develop these medical problems....   [tags: DNA, Gene, Gene expression, Genetics]

Research Papers
878 words (2.5 pages)

Annotated Bibliography On Gene Editing Essay

- Gene editing is one of the most researched fields of molecular biology as manipulated genes provide studying the specific genes and their effects. In the past, there were several attempts to manipulate gene function, including homologous recombination, RNA interference (RNAi), zinc-finger nucleases (ZFNs) and transcription-activator like effector nucleases (TALENs). These past approaches are expensive and time-consuming to engineer as opposed to Crispr Cas9, limiting their widespread use. What the Crispr Cas9 technique offers, and biologists desire, is specificity: the ability to target and study particular DNA sequences in the vast expanse of a genome with ease of preparation as used in Fu...   [tags: DNA, Gene, Gene expression, Genetics]

Research Papers
1153 words (3.3 pages)

Essay on Applications of Protein Purification

- Protein purification has a variety of applications in drug delivery, tissue engineering, and biointerface science. Elastin-like polypeptides can undergo protein purification so that a single type of protein can be isolated from a complex mixture. Because of the increased demands of peptide-based biomaterials for drug delivery and tissue engineering, efficient protein purification techniques must be used. Four techniques that will be discussed in this report are: Affinity chromatography, Centrifugation-based Inverse Transition Cycling (ITC), Microfiltration-based Inverse Transition Cycling, and finally protein purification by self-cleaving purification tags....   [tags: Protein purification, medical, ]

Research Papers
2083 words (6 pages)

Describe How a Gene Encoding a Therapeutic Protein Could Be Cloned into a Vector to Allow Expression

- A vector is a recombinant DNA carrier, all types have three generic properties; introduction to the host cell can be accomplished easily, each vector contains a replication origin enabling it to reproduce inside the cell and in order to determine which cells contain the vector a simple assay can be carried out, such as, growing the cells on agar plates. At present, there are many different types of vectors available for use the best expression system depends on the gene involved (Hartl, 2011). Examples of vectors include; Plasmids, Bacteriophages, Artificial chromosomes, bacteria, cell free systems and viruses (Klug et al., 2003)....   [tags: Vector CLoning, Allow Expression, Recombinant]

Research Papers
1636 words (4.7 pages)

Gene Expression in Eukaryotes Essay

- Distinct characteristics are not only an end result of the DNA sequence but also of the cell’s internal system of expression orchestrated by different proteins and RNAs present at a given time. DNA encodes for many possible characteristics, but different types of RNA aided by specialized proteins sometimes with external signals express the needed genes. Control of gene expression is of vital importance for an eukaryote’s survival such as the ability of switching genes on/off in accordance with the changes in the environment (Campbell and Reece, 2008)....   [tags: DNA Sequence, Nucleotides]

Research Papers
933 words (2.7 pages)

Gene Expression Essay

- Introduction Functional genomics is defined as the use of molecular biological tools to explore the gene functions and interactions from genome sequencing data (SERC, 2013). Expressed Sequence Tag (EST) and Serial Analysis of Gene Expression (SAGE) are among the techniques that are commonly used in functional genomics. The goal of transcriptome studies is to identify the transcripts expressed in a genome. Since human genome studies, EST was the main technique used for transcript identification. Recently, SAGE has been widely used in transcriptome analysis....   [tags: GEnomics, EST, SAGE, transcriptome]

Research Papers
1565 words (4.5 pages)

Gene Expression Noise Essay

- One of the most striking aspects of biology is in the diversity of cellular life. Although much of this variability has been attributed to genetic and environmental factors, recent studies have shown4 that genetically identical organisms in the same environment exhibit heterogeneity in gene expression. This phenomenon, gene expression noise, has been observed and measured across species as divergent as prokaryotes and mammalian cells.1-4 We have been keenly interested in understanding more about the origin of this heterogeneity as well as wondering what potential functional consequences it may have....   [tags: Biology]

Research Papers
748 words (2.1 pages)