Essays on Restriction Enzyme

Introduction

The discovery of restriction enzymes (REs), bacterial transformation, and agarose gel electrophoresis caused the sudden increase of popularity of molecular biology in the 1970s. Restriction enzymes allow the manipulation of specific sites of an organism’s genes and are commonly used in many laboratories worldwide. Restriction enzymes cut DNA in many different ways and can produce sticky ends and blunt ends (Weaver, 2012). The most abundant type of restriction enzymes is Type II, which require magnesium cations as cofactor (Pingoud et al., 2005). Aside from their popularity is recombinant DNA technology as a “cut and paste” tool, estriction enzymes can ...

(Author)

Use of Restriction Enzymes in Pharmaceuticals Restriction enzyme, which is also referred to as restriction endonuclease, is a class of enzyme that is helpful in cutting DNA at restriction sites. Restriction sites are particular recognition nucleotide sequences on double stranded DNA having the length of 4-5 base pairs. Restriction enzymes are also known deoxyribonucleases (DNases). Presently, there are four different types of restriction enzymes: - Type I enzymes that are “classical” in nature, e.g. EcoKI, EcoBI, and EcoR124; - Type II enzymes that are “orthodox” in nature, e.g. EcoRI, HindIII, EcoRV; - Type III enzymes ...

Objective 2

Introduction 2 General background 3 Resistance mutants 4 Hemoglobin mutants 4 Sickle Cell Anemia and Genetics: Background Information 5 Connection of Sickle cell with Malaria 6 Method: 7 Introduction of Recombinant DNA technology in diagnosing disease: 8 Discussion (implementation of recombinant DNA technology to solve the issue) 9 Conclusions 10

References 11

Objective This report targets to study a group of people that has developed resistance to malaria. It will focus on the use of recombinant DNA technology to study this cohort and gain insights into the nature of this observation. This ...

(Author)

Introduction of restriction enzymes Restriction enzymes are highly diverse set of enzymes in bacteria that are used to identify specific 4- to 8-bp sequences referred to as restriction sites and cleave DNA strands internally at this site. These enzymes protect the bacteria from foreign DNA molecules such as those of phages. Restriction enzymes identify specific nucleotide sequences to differentiate between the “domestic” and “foreign” DNA. It attaches to the “recognition site” and breaks both strands of the DNA (Kufe et al.). Restriction enzymes may include EcoRV, EcoRI, and HindIII. EcoRV belongs to the type II restriction/modification system ...

ABSTRACT:

A plasmid DNA is an extra-small circular structure commonly found in bacteria which is capable of replicating by itself in the cell. These are used in genetic engineering as vehicles to carry and make copies of a particular gene which can produce desirable effect. In this report, we discuss the isolation and purification of DNA fromEGFP-LC3 plasmidusing a modified alkaline lysis method and QIAprep spin columns. The concentration of the isolated EGFP-LC3 plasmid DNA was found to be 82.868µg/ml. The ratio of ODs taken at A260nm/A280nm was found to be 5.314 which could be because improper washing step which has ...

Set 1

Part A. 1. BLAST - This algorithm compares the various pieces of primary biological sequence information, such as the nucleotides of DNA sequences or the amino acid sequences of different proteins, in order to identify the library sequences that bear similarities with the query sequence above a certain threshold.

2. ORF Finder – This tool is used when searching for open reading frames (ORFs) in the DNA sequence that the user enters where the information provided includes the protein translation and the range of each open reading frame.

3. Compute pl/Mw tool – This tool computes the isoelectric point (pl) and ...

Lab partners

The transformation of bacterial cells using Puc18 plasmid as a vector Abstract In molecular cloning experiments, the DNA to be cloned is obtained from an organism and then broken into smaller fragments through digestion with restriction enzymes. The fragments of restriction enzyme digestion are then ligated /combined with some vector DNA to produce recombinant DNA molecules. These recombinant DNA molecules are then introduced into a host organism. When the host organism replicates, the recombinant molecule is replicated together with it. With time, large populations of transformed cells or clones are generated expressing the genotype and phenotype of the inserted ...

Determining the size of gene fragments

1) Alul restriction enzyme cuts the protease gene at restriction sites with base position 62 and position 1149 respectively. Considering the fact that the gene has 1216 base pairs, three possible fragments are generated with Alul restriction digestion.

The first digestion fragment length ranges from base pair 1 to base pair 62= 61 base pairs.

The second fragment length ranges from base pair 62 to base pair 1149 in length=1087 base pairs The third fragment ranges from base pair 1216 to base pair 1149 in length= 67 base pairs.

2) Sa/l restriction enzyme cuts the protease gene at ...

Introduction

The article, “Genotyping and identification of mycobacteria by fingerprinting Techniques” by Khosravi and Seghatoleslami (2009) shows how fingerprinting can be used to identify mycobacteria in combating Tuberculosis (TB) which for a long time has been a serious health concern by the World Health Organization (WHO). Some of the techniques used in molecular epidemiology like MIRU-VNTR and spoligotyping are described in the article.

Summary

The article has deeply illustrated the various ways of carrying out Genotyping of mycobacteria for carrying out epidemiological studies. It has also shown that through molecular typing of MTB the disease can be controlled. The study of the epidemiology ...