Partial cloning of prohibitin cDNA from canine, feline, bovine, equine, and rabbit liver mRNA by RT-PCR.

This research explores the partial cloning of the prohibitin gene from different animal liver cells, using a technique called RT-PCR, and analyzes the similarities among these cloned sequences.

Cloning of Prohibitin cDNAs

• The researchers focused on a protein known as prohibitin, which acts as an inhibitor to cell growth, and its gene is potentially a tumor suppressor gene.
• They performed partial cloning of prohibitin complementary DNAs (cDNAs) from the liver messenger RNAs (mRNAs) of various animals – canine, feline, bovine, equine, and rabbit – using Reverse Transcription Polymerase Chain Reaction (RT-PCR).
• RT-PCR is a popular molecular biology technique used to copy and amplify specific DNA sequences from an RNA template. It first converts RNA into cDNA through reverse transcription, and then amplifies it using PCR.

Homology Analysis of Cloned Prohibitin cDNAs

• The next step involved the comparison of the sequences of these cloned prohibitin cDNAs from different animals with each other, and with the prohibitin sequences reported for human and rat.
• This was done to evaluate the homology or similarity among these sequences.

Results and Conclusion

• The homology or sequence similarity in the prohibitin cDNA region among the different animals was found to be approximately 90%.
• Furthermore, the amino acid sequences of each RT-PCR product shared more than 95% identity, indicating that these sequences were very similar in their structure and composition.
• Based on these findings, the researchers concluded that all the RT-PCR products are a part of the prohibitin cDNA of each animal.
• This high degree of sequence similarity suggests that the prohibitin gene has been highly conserved across different species, further supporting its crucial role in cell growth and potentially in tumor suppression.