Volume 1, Issue 1 (2010)                   JMBS 2010, 1(1): 0-0 | Back to browse issues page

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Esmaeilzadeh Khorasani M, saadati M, Agaei pour K. Separating and Cloning fusion gene of measles virus (AIK-C) vaccine strain. JMBS 2010; 1 (1)
URL: http://biot.modares.ac.ir/article-22-2371-en.html
1- Tehran
2- Imam Houssein University, Tehran
3- tehran
Abstract:   (16726 Views)
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.
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Received: 2010/07/22 | Accepted: 2010/11/15 | Published: 2010/12/13

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