Abstract
Introduction: Measles virus (MV) belongs to the morbilivirus genus of paramyxoviridae family, and has single stranded, negative polarity, non segmented RNA genome.
Method: In this research, the total RNA was extracted of measles virus (AIK-C) vaccine strain. The extracted RNA was immediately used in reverse transcription reaction to generate cDNA. The 1st strand cDNA was used to amplify the F gene by specific primers in a reaction PCR. The PCR product with the expected size of 1662 bp was cloned into expression plasmids pET-22b(+) and pET-28a(+).
The recombinant plasmids were transformed into competent E.coli DH5α cells and clonies were screened with direct PCR. The recombinant plasmids were extracted by Alkaline lysis and were compared with non- recombinant plasmids in molecular weight.
Results: Recombinant plasmids were digested with Nde I and Hind III restriction enzymes. The DNA band with an approximate size of 1662 bp was detected on 1.5% agarose gel. The recombinant plasmid pET-28a(+) was sequenced, comparison of this sequence with the coding sequence F protein of measles virus (AIK-C) in Genbank (AF266286) was revealed high degree of homology and showed that F gene is highly conserved.
Conclusion: It was showed that F gene is highly conserved. Thus F gene is important for studing in order to produce recombinant vaccine.