ساخت وکتور ریپورتر القایی لوسیفراز برای ردیابی فعالیت ویروس HTLV-1

 Background: HTLV-1 (Human T_cell lymphotropic virus type), with about 20 million individuals infected worldwide, is a global health problem and is endemic in certain area such as Japan and Khorasan province of Iran. HTLV-1 is the causative agent of progressive diseases, Adult T cell Leukemia (ATL), and HTLV-I Associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP) which have no yet approved effective treatment. Due to non cytolytic characteristic of this virus and the cell associated properties, there is no routine producer for drug study in vitro and its confined to some HTLV infected cell lines. Therefore construction of a reporter vector is necessary to evaluate HTLV-1 infectivity in cell culture for drug studies. Designed reporters for retroviruses are usually based on LTR transactivatin. LTR region of HTLV-1 contains virus promoter that plays the most important role in replication and transcription by Tax transactivation effect. Methods: LTR region was digested from pUCLTR-Lacz by HindIII and subcloned into MSC region of a promoterless reporter vector, pGL4.17, upstream to luciferase gene. Colonies were screened by Colony PCR, then selected colonies were confirmed by RE digestion and sequencing. HEK 293T cell line was transfected by recombinant vector and inducibility expressing of luciferase was evaluated. Results: Recombinant vector has expressing levels more than 50 folds compared to control when co-transfected with Tax expressing vector into HEK293T cells. Conclusions: According to high functionality of produced recombinant vector, it seems a good applicable tool to make an indicator cell line in subsequent basic and drug studies.