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Iranian Journal of Microbiology، جلد ۷، شماره ۴، صفحات ۲۲۱-۲۲۵
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عنوان انگلیسی Designing a recombinant Bacmid construct of HCV core+1 in Baculovirus expression system
چکیده انگلیسی مقاله Background and Objectives: Hepatitis C virus (HCV) chronically infects around 200 million people worldwide and fre- quently causes liver cirrhosis and hepatocellular carcinoma. Rapid detection of this virus results in decreasing the distance between infection and initiation the anti-viral treatment, and may prevent most of the undesirable consequences. The new detected HCV protein "Core+1" made from the ribosomal frame shift in Core region is an important candidate for diagnostic tools. This study was conducted to design a recombinant Bacmid plasmid expressing the HCV 1a Core+1 sequence in the Baculovirus expression system for further diagnostic applications. Materials and Methods: The HCV Core +1 gene was amplified by PCR using the pcDNA-HAF recombinant vector that contained the Core+1 sequence from HCV genotype 1a as a template, and the specific primers with 2 restriction sites for Nco I and Xba I restriction enzymes. The PCR product was cloned in XbaI/NcoI restriction sites of the linearized p FastBac-HTB vector and evaluated by using those restriction enzymes and sequencing. Then the recombinant p FastBac-HTB vector was transformed in DH10Bac and the result was screened and confirmed by X-Gal discrimination and PCR. Results: The HCV 1a Core+1 was successfully amplified and the PCR product was confirmed by using the related restriction enzymes and sequencing. Cloning of p FastBac vector with the purified PCR product of HCV Core+1 was confirmed. Finally, the recombinant Bacmid was successfully transformed in DH10Bac. Conclusion: The recombinant Bac-Core+1 expression vector is considered as an important tool to transfect the sf9 cell line and expression the Core+1 protein.
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پونه رحیمی | pooneh rahimi
department of hepatitis and aids, pasteur institute of iran.

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

محمدرضا خاتمی نژاد | mohammadreza khataminejad
department of biology, islamic azad university, tonekabon branch, tonekabon, ir iran.

سازمان اصلی تایید شده: دانشگاه آزاد اسلامی علوم و تحقیقات (Islamic azad university science and research branch)

نشانی اینترنتی http://ijm.tums.ac.ir/index.php/ijm/article/view/786
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