Apolipoprotein E Gene Expression in the Liver of Genetically Obese Zucker Rats:

Apolipoprotein (apo) E is a protein involved in both lipid metabolism and neuroprotection. Recently, it has been suggested that apoE may play a role in the regulation of food intake and body weight in rodents. However, rodent plasma apoE is difficult to purify in reasonable amounts due to numerous time-consuming steps. To circumvent this, we created a bacterial expression system for the efficient production of large amounts of rat apoE. We inserted rat apoE DNA into the pET30 expression vector and overexpressed the proteins in Escherichia coli strain BL21 (DE3). A histidine tag present at the N-terminus allowed for easy purification of the recombinant protein. The tag was removed with an IgA protease (Igase) from Neisseria gonorrhoeae leaving the mature form of the protein. The use of Igase was important as several more common proteases routinely cleave apolipoproteins at undesired sites. The recombinant protein was then compared both structurally and functionally to rat plasma apoE. This expression system will be highly useful for probing the ability of rat apoE to mediate food intake in rats.

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Dietary fat up-regulates the apolipoprotein E mRNA level in the Zucker lean rat brain

Knight

Mahajan

Qiao

2001

Neuroreport

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High-fat diet alters apo E-dependent processing of beta-amyloid precursor protein. Here we have evaluated the effects of dietary fat on brain apo E mRNA in Zucker lean and obese rats. After approximately 2 months on a high-fat diet, there was significant up-regulation of brain apo E mRNA in the Zucker lean rat in parallel with weight gain. Densitometric quantification revealed a 17% increase in apo E mRNA in the brains of lean rats fed high-fat diet compared with those of lean rats fed rat chow. No significant difference in brain apo E mRNA of Zucker obese rats fed different diets was found. These results suggest that dietary fat alters brain apo E levels, which may be regulated, in part, through the leptin receptor.

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Apolipoprotein E Gene Expression in the Liver of Genetically Obese Zucker Rats:

Cited by 3 publications

References 8 publications

Plasma lipoproteins and apolipoproteins B and E in rats fed a high-fiber fructose-based diet

Plasma lipoproteins and apolipoproteins B and E in rats fed a high-fiber fructose-based diet

Bacterial expression and characterization of rat apolipoprotein E

Dietary fat up-regulates the apolipoprotein E mRNA level in the Zucker lean rat brain

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