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Summarizing a biological article - Essay Example

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The E.coli cells used had shown overexpression of inaZ gene that codes ice nucleation protein. Besides purification of active ice nucleation protein, the study also aimed at…
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Summarizing a biological article
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Phospholipid Analysis and Fractional Reconstitution of the Ice Nucleation Protein Activity Purified from Escherichia coli Overexpressing the
inaZ Gene of Pseudomonas syringae
The study aimed at purification of active ice nucleation protein from recombinant E.coli cells membranes. The E.coli cells used had shown overexpression of inaZ gene that codes ice nucleation protein. Besides purification of active ice nucleation protein, the study also aimed at demonstrating that it is possible to restore activity of ice nucleation protein after reconstitution procedures of mostly dilapidated proteins. The report also presented and analyzed data on the contents of phospholipids obtained from fractionation of E.coli cells proteins that showed ice nucleation activity (Palaiomylitou et al. 67).
The study addressed initial questions relating to the role of membrane phospholipids in ice nucleation activity. Previous studies have also tried to address role of phospholipids as a factor of ice nucleation activity in most bacterial membranes. The study established the role of phospholipid components such as phosphatidylinositol and phosphatidylethanolamine, to ice nucleation activity.
The researchers in the study grew E.coli bacterial strains in controlled cultures at 21 degrees Celsius and pH of 7. The study monitored the growth of the cells through measurements of absorbance of the media at 600nm using Perkin-Elmer spectrophotometer. The researchers the harvested multiplied cells after the end of logarithmic phase. The study then overexpressed ice nucleation proteins E.coli cells through cloning in Pt3t7lac vector. The study assayed Ice nucleation activity in the cells through droplet-freezing assay followed by serial dilutions. The study finally used ICE software program to calculate ice nucleation activity. The study then extracted ice nucleation protein through lysis of the E.coli cells and the protein content determined through Bradford method. The researchers then purified and analyzed Ice nucleation protein through gel electrophoresis followed by immunoblotting (Palaiomylitou et al. 68). After production of protein antibodies, the study extracted the lipids through monophasic system involving methanol, chloroform, and water ratio of 2:1:0.8.
The study affirmed the restoration effect of phospholipids on the activity of ice nucleation protein after the delipidation of lipoproteins. However, must phasmatoscopic analysis to help in the elucidation of hydrophobic properties of ice nucleation protein and to determine the mode of protein-lipid interaction in the ice nucleation protein (Palaiomylitou et al. 75).
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Work Cited
Palaiomylitou, M. A., Kalimanis, A., Koukkou, A.I., Drainas, C., Anastassopoulos, E., Panopoulos, N.J, Ekateriniadou, L. V. and Kyriakidis, D. A. “Phospholipid Analysis and Fractional Reconstitution of the Ice Nucleation Protein Activity Purified from Escherichia Coli Overexpressing the inaZ Gene of Pseudomonas Syringae.” Cryobiology 37.1 (1998): 67–76. Read More
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