Laboratory Report on Gene Cloning into Expression Vector
Uploaded by CaseyP on Jun 14, 2017
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Laboratory Report on Gene Cloning into Expression Vector
Abstract
Gene cloning involves manipulation of a given cellular component to isolate the desired gene properties. Isolation of particular cellular component forms the core of the cell chemistry and biology. For a gene to be efficiently cloned, it must be inculcated in a larger medium that will allow sufficient expression of its markers. The vectors form the structures viable enough to encourage the manipulation of the genes to get the desired efficacy. Mostly used vectors are the plasmid vectors. They are ideal because of their neutrality nature and the growth favoring characteristics. The cloning vector obtained from the plasmid is useful and provides a good binding for the foreign DNA fragments which allows for the elucidation of cloned gene. This laboratory report explores on how gene cloning is altered into an expression vector. It also highlights on the Polymerase Chain Reaction and how the process is used to amplify the cloned genes into the desired particles. The last section explains the findings of the experiment.
Keywords: cloning vectors
Polymerase Chain Reaction
Introduction
For any desired effect of gene cloning to have an impact, there must be a proper layout on the ways to elucidate the genes properly. The use of expression vector is effective method since the vectors provide the necessary requirements and conducive media for an insertion of foreign DNA genes. In the experiment, tomato cells were used as the expression vector to plant with the gene of interest. Ideally, for a successful experiment to be conducted on the gene cloning, different sets of parameters must be taken into consideration. First, the gene has to be isolated. Secondly, the gene has to be cloned into a vector for a greater expression, and lastly, the gene must be expressed and translated to give the desired aims of the experiment (Brown 10). The laboratory experiment conducted followed the same fashion. Adjustments were made in the vector to meet the typical qualities of eukaryotic cells. The first step taken into consideration was the use of promoter sections, Kozak sequence and inserting appropriate start and stop codons. In addition to the requirements, kanamycin resistance gene was used as selected marker to identify the cells that have taken up the DNA.
Materials and Reagents Used
Isolated cDNA gene
Plasmid, pXCN
Escherichia coli strain
5microlitres buffer
3microlitres...