Tuberculosis and Respiratory Diseases
- Volume 39 Issue 3
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- Pages.242-247
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- 1992
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- 1738-3536(pISSN)
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- 2005-6184(eISSN)
Quantitative Measurement of Surfactant Protein B mRNA by Filter Hybridization
Filter Hybridization 방법에 의한 Surfactant Protein B mRNA의 정량측정
- Park, Sung-Soo (Department of Internal Medicine, Hanyang University College of Medicine) ;
- Lee, Dong-Hoo (Department of Internal Medicine, Hanyang University College of Medicine) ;
- Shin, Dong-Ho (Department of Internal Medicine, Hanyang University College of Medicine) ;
- Lee, Jung-Hee (Department of Internal Medicine, Hanyang University College of Medicine)
- Published : 1992.06.30
Abstract
Background: The ability to precisely measure specific mRNA levels by hybridization to complementary DNA probes is an important tool for analyzing the regulation of gene expression. Surfactant proteins have important roles in regulating surfactant metabolism as well as in determing its physical properties. Method: The complete coding regions for rat surfactant protein complementary DNA of surfactant protein B were subcloned into pGem 3Z or 4Z such that either antisense or sense transcripts were obtained by using SP 6 RNA polymerase. Surfactant protein B mRNA was measured by filter hybridization. Results: Equation of standard curve between counts per minute (Y) and surfactant protein B mRNA transcript input (X) was Y=2034.9 X+159.1. Correlation coefficient was 1.0. Couclusions: Filter hybridization assay is suited to situation when rapid, accurate quantitation of multiple samples is required.
연구배경 : Surfactant 단백은 surfactant의 물리학적 성상의 결정 및 대사를 조절하는데 있어서 중요한 역할을 한다. 유전자 발현의 조절을 연구하기 위하여서는 cDNA의 탐지자에 의한 mRNA의 정량측정이 중요하다. 방법 : 쥐의 surfactant 단백 B의 cDNA에 대한 coding 부위를 PGem 3Z 또는 4Z에 subclone하여 SP6 RNA polymerase 효소를 이용하여 antisense와 sense을 얻었다. Sense을 이용한 filter hybridization올 시행하여 정상곡선을 얻었다. Antisense는