PCR AMPLIFICATION AND ANALYSIS OF VARIATION IN GROWTH HORMONE GENE OF CETARTIODACTYLA

Abstract

Background: The sequence of growth hormones (GHs) is generally conserved in mammals but short bursts of rapid changes occur during evolution of primates and artiodactyla. This has led to marked differences in primary structure and biological specificity in cetartiodactyla. Present study was done to amplify and further explore GH evolution in cetartiodactyla. Methods: Buffalo placental tissue was collected from Lahore, Pakistan and stored at -20?C. Placenta was homogenized and RNA was extracted and analyzed on horizontal agarose gel electrophoresis. RT-PCR (reverse transcription polymerase chain reaction) was performed by using oligo(dT) primers. GH gene was amplified by PCR and analyzed on agarose gel electrophoresis. PCR product was purified and sent for sequencing. Sequence alignment was done using bioinformatics tools. Results: GH gene was successfully amplified. Silent mutations in the sequence of Bubalus bubalis GH were compared with the reported sequence of bovine GH and variations were found from reference sequence. Conclusion: In conclusion, GH gene has variation in sequence during evolution due to different factors. A total of 10 variations were observed in the forms of transitions, transversions and deletion as compared to reference sequence.