Quantitative measurement of surfactant protein B mRNA by filter hybridization. |
Sung Soo Park, Dong Hoo Lee, Dong Ho Shin, Jung Hee Lee |
Department of Internal Medicine, Hanyang University Collage of Medicine, Seoul, Korea |
|
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 determining its physical properties.
Methods The complete coding regions for rat surfactant protein complementary DNA of surfactant protein B were subcloned into pGem 32 or 42 such that either antisense or sense transcripts were obtained by using SP 6 RN A polymerase. Surfactant protein B mRNA was measured by filter hybridization.
Results Equation of standard cuπe 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.
Conclusions Filter hybridization assay is suited to situations when rapid, accurate quantitation of multiple samples is required |
Key Words:
Filter hybridization, Quantitation mRNA, Surfactant Protein B |
|