Essay Sample on Cloning the Genes of a Mouse

Goal 1: Cloning the Gene

To clone the genes of a mouse, one will have to follow the following procedures. First of all, identify a vector which has small molecules which are easy to isolate and characterize the intended gene we want to clone. The procedure for cloning will depend on different organisms; thus one should have explicit knowledge of the organism's characteristics to begin the process of isolating a gene and cloning it. One has to consider an appropriate vector to start the work. The vectors with smaller molecules tend to be easy to isolate and have to be able to replicate within the cell to allow the alien donor fragment can be easily amplified within the cell. Another consideration factor is that one has to locate a site which is restricted so that the insertion of the Deoxyribonucleic acid (DNA) can take place (Griffiths, 1999). It is advised for one to find a site which can be inserted in one side of the vector. Also, there has to be a unique way to identify and recover the combined molecule.

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For a more substantial part of the process, to clone a gene one has to have extensive knowledge of the gene and the procedure beginnings with the analysis of the eukaryotic genomic DNA and followed by extraction of a more substantial part of the clone obtained from the DNA sample. The collected samples are what is known as the DNA library and provide information about the specified DNA and its properties (Griffiths, 1999). The following work is to find the DNA clone. One should note that the different DNA vectors usually have varying amounts of DNA; thus the chosen DNA vector for the library will depend on the size of the genome which will be made into the library.

The second factor to consider is whether one will need to make a genomic library or a cDNA library which is a complementary DNA, a synthetic DNA constructed from the mRNA from an enzyme called reverse transcriptase which is derived initially from retroviruses (Griffiths, 1999). While using mRNA s a template in the cloning of the gene, the reverse transcriptase is used to make as a single-thread molecule of the DNA that can be used to synthesize the DNA molecule, and it is not void of the downstream and the upstream of the molecules regulatory sequential and intones and in most cases one has to limit the genomic factors in the library as it is helpful when the person doing the cloning of a mouse gene to know what chromosomes are contained in the gene. In this process, one has to use the well-known procedure to conduct the genetic cloning of mammals and this done by sorting the genetic chromosomes of the mammal which in this case is the mouse provided for the procedure using an instrument called flow cytometer. This tool is used by passing a stream of chromosomes through it, and the apparatus works by sorting these chromosomes according to their size. After the chromosomes are sorted, the chromosomes which are required for the procedure are then collected and used to clone the genetic components of the DNA of the mouse.

Goal 2: Characterizing Tissue Expression

If the mouse provided is noted to have small chromosomes, there is another procedure which can be used to help in cloning the genes of the mouse, and it is called pulse gel field electrophoresis. This well-known technique is used to fractionate the nucleic acid or the specified protein when it is put under the influence of an electric field which is strong. The Rab proteins are a subfamily of the ras family and they are low molecular weight GTPases which is involved in the transportation of intracellular vesicular and regulation.

Currently this report is indicating the cloning and characterization of the mouse homologue of Rab32. It indicates the murine Rab32 shows a ubiquitous expression pattern, with tissue specification in variation expression level. It is noted that there are three types of cells having specialized organelles and have high rate of Rab32 that is the platelets, mast cells and melanocytes.

Goal 3: Introducing Mutation Into the Cloned Gene

There is a discovered method of introducing the mutation into the cloned gene and the method is called the polymerase chain reaction (PCR) and it is widely used for site-directed mutagenesis. There has to be a suitable restriction site in the scene of the mutated nucleotide is needed so that recloning can occur because introduction of mutation can only be done on the primer sequence used for PCR. There are several methods that have been put to place to limit this disadvantage. For the meantime the method of overlap of the extension method is used (Ho et al., Gene 77 (1989) 51-57) as it has a wide advantage over the previously used methods. Three primers and a series of primers specifically used for various mutations are synthesized chemically. When the correct oligodeoxyribonucleotides are picked as common primers, every mutation will require one additionally primer only. This method is thus very vital in introducing various mutations in many sites of the targeted DNA which in this case is the mouse'.

Goal 4: How to Knock a Mouse

Knocking up a mouse is one of the greatest achievements a researcher can accomplish in the issue of genetics and mutation. There are various ways which one can use to generate a knock-in mouse and all these ways have one purpose and that is to introduce exogenous genes which have a potential to alter the state of endogenous genes and so as to generate some kind of change in the body of a mouse at a genetic level. One of the feature to generate a knocked up mouse is to use a convection knock-in mouse where a single or multiple mutation points are targeted or engineered the gene of the mouse which is in interest with an insertion of a cassette to illustrate a sequence which is alternate to the original. The process of generating a mouse knock-in follows the same procedure as that of producing a knock out mouse and this goal is by applying the use of homologous recombination.

Reference

Griffiths, A. J. (1999, January 01). Cloning a Specific Gene. Retrieved from https://www.ncbi.nlm.nih.gov/books/NBK21450/