نویسندگان
1 مرکز تحقیقات بیوتکنولوژِی، انستیتو پاستور ایران
2 گروه میکروبشناسی، دانشکده پزشکی، دانشگاه شاهد
3 انستیتو پاستور ایران
چکیده
کلیدواژهها
عنوان مقاله [English]
نویسندگان [English]
Background and Objective: The aim of the present study was
to express recombinant human interferon-gamma in Leishmania tarentolae.
The Leishmania expression system represents the combination of easy handling
known from bacterial expression systems with the potential of a eukaryotic
protein expression, folding and modification system. The trypanosomatid
protozoan host Leishmania tarentolae was isolated from lizard and is not
pathogenic to mammals. Materials and Methods: In this study, two constructs were
developed for expression of recombinant human interferon-gamma (hrIFNg) in L.
tarentolae. Each one of constructs carries an antibiotic resistant gene
such as Hygromycin and Nourseothericin. For high level expression of the human interferon-gamma, developed DNA
cassette that contains interferon-gamma (IFNg) cDNA was designed to integrate into
a genomic small subunit rRNA locus of L. tarentolae by homologous
recombination. Results: The integration of the expression cassette into
the ssu locus was confirmed by diagnostic PCR of the genomic DNA of transgenic
strain. Conclusion: Although it has been shown that E. coli
might be considered as a suitable host with a relatively high level of
expression and lack of posttranslational modifications and the need for
refolding stages are still big challenges. In contrast, L. tarentolae is
eukaryotic gene expression machinery, which includes full glycosylation and
disulfide bond formation, and thus represents a potential advantage over other
expression systems. These facts confirm that the gene expression system using
Leishmania parasites combines many of the advantages of both prokaryotic and
eukaryotic expression systems.
کلیدواژهها [English]
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