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Title 

Differentially expressed aortic genes in cholesterol-fed rabbits

Authors 

Tae Gyun KimMoon Kyoo JangNam Ho JeoungMyung Sook ChoiSong Hae BokJu Won KwakYong Bok Park

Publisher 

Springer Verlag (Germany)

Issue Date 

1998

Citation 

Molecules and Cells, vol. 8, no. 3, pp. 324-329

Keywords 

AtherosclerosisAtherosclerotic LesionCholesterolDDRT-PCR

Abstract 

In order to identify key genes involved in the development of atherosclerotic lesions, differentially expressed genes in atherosclerotic plaques obtained from diet-induced hypercholesterolemic rabbit aorta were screened using the differential display (DD) RT-PCR technique. Aortic RNAs were isolated from rabbits fed cholesterol-supplemented (2% cholesterol in lab-chow, w/w) chow diet for 12 weeks, followed by the synthesis of cDNAs by reverse-transcription using 2-base anchored oligo(dT) (5′-T11VN) as 3′-primers. Synthesized cDNAs were amplified by PCR using arbitrary 10-mers as 5′-primers and the same 3′-primers used in the reverse-transcription. Amplified cDNAs sized between 0.2 to 0.5 kb obtained from control and cholesterol-fed rabbit aortas were displayed on the 6% DNA-sequencing gel for comparisons. The cDNA bands showing distinctive differences in patterns of display or in density of the band were extracted from the gel. A total of 66 differentially displayed cDNAs was isolated and subjected to the reverse-Northern and Northern blot analyses in order to confirm the differences. Through the extensive confirming processes, three cDNAs were finally selected (designated CRGRA-1 through -3) and their nucleotide sequences were determined. Two of those (CRGRA-1 and -2) were determined to be up-regulated and the other (CRGRA-3) was down-regulated by the cholesterol-feeding. Upon homology search on databases for the identification of the genes, the first cDNA (CRGRA-1) turned out to be a part of a novel gene, the second one (CRGRA-2) was homologous (82%) to the corresponding segment of mitochondrial NADH dehydrogenase subunit 4 gene, and the last one (CRGRA-3) was identified to be homologous (94%) to a segment of human small GTP-binding protein (Rab7) gene.

ISSN 

1016-8478

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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