Locate an article entitled \"A simplified method for reconstituting active E .co

Question

Locate an article entitled "A simplified method for reconstituting active E .coli DNA polymerase III" in the journal of Protein and cell. It was published in 2011 and spans pages 303-307.The first Author of the article is Shi-Qiang Lin.Read the article and answer questions below:

a) What was the overall aim of the research conducted? (3marks )

b)In the article the author cite many other publications. Each citation is associated with corresponding fact mentioned by the authors. Choose one fact listed by authors and locate the associated publication that is cited.Then go and confirm if the fact can be found in that publication. Explain how you located the other publication as well as how you confirmed that the information listed in the article by Pich and colleagues is correct.(8 marks )

c) List and briefly describe two techniques or experimental approaches the researchers used that are also covered in your study material for this unit. State what the researchers intended to achieve with each technique (e.g X-Ray diffraction to get the structure of protein X)(10 marks)

d)Provide a citation for the article as well as full reference (4 marks )

Explanation / Answer

a. The overall goal of the research conducted is to coexpress and purify both DNA pol III core complex and clamp loader complex in a very elegant way that harness the protein-protein interaction of the subunits involved to balance the stoichiometry needed for reconstructing in-vitro DNA polymerization activity.

b. The authors here cite a very versatile protein coexpression strategy which was published in a very reputed journal "Nature methods" byy Tolia and Joshua in 2006. Now a days its very easy to locate an article in the internet. We can simply google with the title of the paper and click on the first relevant hit or we if we know the DOI associated with the paper we can directly locate the paper from Pubmed central. After reading the cited paper one can be more evident with the facts or the techniques used in that paper which will help comprehending the current article with ease.

c. 1. Ni-NTA affinity chromatography for Protein Purification:They expressed the protein with a histidine tag which is needed for the purification of the using a choratographic column with intact Ni-NTA.

2. Gap Filling experiment: This experiment was performed to analyse or determine the activity of the purified DNA pol III complex in vitro.

d. Lin, Shi-Qiang, Li-Jun Bi, and Xian-En Zhang. "A simplified method for reconstituting active E. coli DNA polymerase III." Protein & cell 2, no. 4 (2011): 303-307.

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