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KMID : 0364219900330010035
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Korean Journal of Zoology
1990 Volume.33 No. 1 p.35 ~ p.44
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A Study on the Molecular Biology of Renin-Angiotensin System : Renin Gene Expression and Construction of Genomic Library
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Park Young-Soon
Han Dong-Min
Kim Chong-Ho
Moon Young-Hoe
Lee Ho-Sub
Ko Geon-Il
Kim Sung-Joon
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Abstract
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Poly (A) + RNA was isolated from mouse submaxillary gland and renin mRNA was isolated by poly (U)-sepharose chromatography and sucrose linear densiW gradient centifugation. And renin mRNA was identified by in vitro translation and immunoprecipitation. In order to construct recombinant plasmid, renin cDNA was synthesized by using reverse transcriptase and inserted into EcoRi site of PUC19. In addition, the cDNA was also synthesized using polymerase chain reaction and inserted into HindlIl site of PUC19. The recombinant plasmid was transformed into JMlO3 and the expression of the inserted renin cDNA was examined. The transformant produced renin protein having a molecular weight of 45, 000 dolton, which showed hypertensive effect upon injecting it into rabbit ear vein. A renin genomic library was prepared by inserting rabbit kidney DNA into EMBL3 phage, and was screeined for the isolation of renin gemomic DNA using renin cDNA probe.
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KEYWORD
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Submaxillary gland renin , Kidney renin , Renin cDNA
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