Teimourpour Roghayeh, Zare Hosna, Rajabnia Ramazan, Yahyapour Yousef, Meshkat Zahra
Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran.
Antimicrobial Resistance Research Center, Mashhad University of Medical Sciences, Mashhad, Iran.
Iran J Microbiol. 2016 Apr;8(2):132-8.
HBHA and Mtb32C have been isolated from culture supernatants of Mycobacterium tuberculosis (M. tuberculosis) and Mycobacterium bovis (M. bovis) and their immunogenicity previously studies have been confirmed. In this study, capability of constructed vector containing two mycobacterial immunodaminant antigens (Mtb32C-HBHA), in producing new chimeric protein under the in-vitro condition was examined.
In present study Huh7.5 cells was transfected with Mtb32C-HBHA -pCDNA3.1+ recombinant vector using the calcium phosphate method and expression of chimeric protein was assessed by RT-PCR and Western blot methods.
Results of RT-PCR and Western blot showed expression of 35.5 KD recombinant protein (Mtb32C-HBHA) in this cell line.
The constructed vector can produce two highly immunogenic antigens that fusion of them to gather makes chimeric antigen with new traits. Other attempts are needed to evaluate specific properties of this new antigen such as molecular conformation modeling and immunologic characteristics in future studies.
HBHA和Mtb32C已从结核分枝杆菌（M. tuberculosis）和牛分枝杆菌（M. bovis）的培养上清液中分离出来，其免疫原性先前已得到证实。在本研究中，检测了包含两种分枝杆菌免疫优势抗原（Mtb32C-HBHA）的构建载体在体外条件下产生新嵌合蛋白的能力。
在本研究中，采用磷酸钙法用Mtb32C-HBHA -pCDNA3.1+重组载体转染Huh7.细胞，并通过RT-PCR和Western印迹法评估嵌合蛋白的表达。
RT-PCR和Western印迹结果显示在该细胞系中表达了35.5 KD重组蛋白（Mtb32C-HBHA）。
构建的载体可产生两种高免疫原性抗原，它们融合在一起形成具有新特性的嵌合抗原。在未来的研究中，需要进行其他尝试来评估这种新抗原的特定特性，如分子构象建模和免疫特性。
