cloning and expression of rabies virus glycoprotein gene into eukaryotic system

Authors

k borhani department of virology, school of medical sciences, tarbiat modares university, tehran, iran

t bamdad department of virology, school of medical sciences, tarbiat modares university, tehran, iran

m ajorloo department of virology, school of medical sciences, tarbiat modares university, tehran, iran

shr mozhgani human rabies vaccine laboratory, production and research complex pasteur, institute of iran, tehran, iran

abstract

background and aims: the aim of this study was cloning and expression of rabies virus glycoprotein by a eukaryotic expression plasmid pcdna3.1(+) in bsr cell line. this construct might be used for a potential dna vaccine. materials and methods: glycoprotein gene was synthesized and cloned into pbluescript vector and then sub cloned into eukaryotic expression vector (pcdna3.1(+)). after verification of the cloning, the recombinant plasmid was transfected into bsr cell line (a clone of bhk-21 cell), and its expression was detected by rt-pcr. results: the authenticity of the recombinant plasmid pcdna3.1(+)-gp has been confirmed by a quick check method and restriction endonuclease digestion analysis, and after transfection into eukaryotic cells, the presence of mrna transcripts was verified by reverse transcriptase-polymerase chain reaction (rt-pcr). conclusion: this study demonstrated that the construction of eukaryotic expression plasmid for rabies virus glycoprotein is possible. nevertheless, more work is necessary to develop this kind of vaccine for final use.

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Journal title:
iranian journal of virology

جلد ۶، شماره ۲، صفحات ۸-۱۱

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