Cloning of Heart-type Fatty Acid-binding Protein (H-FABP ) Gene of Swine and Difference of Its Expression at Different Stages
Huang Yu-hai
Abstract
Huang Yu-hai
Abstract
Heart-type fatty acid-binding protein(H-FABP) gene mRNA extracted from muscle of pig(Duroc×Landrace×Yorkshire) was amplified using RT-PCR. And a DNA fragment of 211 bp in length was obtained and the PCR product was cloned into PGEM-T vector. The H-FABP gene was isolated and sequenced from the positive clones screened. Sequence analysis suggested that this fragment was partial sequence of H-FABP cDNA. The gene homology of fragment obtained in this study compared with that of reported H-FABP sequence in muscle of porcine was 99%. Based on the H-FABP gene clone, an optimal semi-quantitative RT-PCR method was successfully constructed. Using 18S rRNA as inner control, the difference of H-FABP gene expression at different stages of swine was researched. The result indicated that the H-FABP gene expression level decreased from 1 to 50 kg, and increased from 50 to 90 kg of body weight.
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Heart-type fatty acid-binding protein(H-FABP) gene mRNA extracted from muscle of pig(Duroc×Landrace×Yorkshire) was amplified using RT-PCR. And a DNA fragment of 211 bp in length was obtained and the PCR product was cloned into PGEM-T vector. The H-FABP gene was isolated and sequenced from the positive clones screened. Sequence analysis suggested that this fragment was partial sequence of H-FABP cDNA. The gene homology of fragment obtained in this study compared with that of reported H-FABP sequence in muscle of porcine was 99%. Based on the H-FABP gene clone, an optimal semi-quantitative RT-PCR method was successfully constructed. Using 18S rRNA as inner control, the difference of H-FABP gene expression at different stages of swine was researched. The result indicated that the H-FABP gene expression level decreased from 1 to 50 kg, and increased from 50 to 90 kg of body weight.
Key concepts: Fatty acid-binding protein, Heart-type fatty acid binding protein, Biology, Gene, Cloning (programming), Molecular biology, Complementary DNA, Sequence analysis