Isolation, cloning and molecular analysis of ag85a and tb10.4 genes from Mycobacterium tuberculosis

Background: Novel tuberculosis (TB) vaccines that aim to boost and/or replace Bacillus Calmette-Guerin (BCG) are currently in development. DNA vaccines can stimulate both humoral and cell-mediated immunity in different animal models of TB and is thought to be a promising strategy in the development of new vaccines against TB. The aim of this study was to design and construct a DNA vaccine encoding ag85a and tb10.4 fusion genes of Mycobacterium tuberculosis.Materials and Methods: tb10.4 fragment was amplified by PCR and the product was digested with restrictionenzymes. Next, it was cloned into the pcDNA3.1+ plasmid. The ag85a gene and pcDNA3.1+/tb10.4 plasmid were digested by EcoRI and BamHI restriction enzymes. Constructed vector was sequenced. The molecular analysis was done using bioinformatics software. New chimeric vector containing ag85a-tb10.4 genes were purified. Expression of pcDNA3.1+/tb10.4-ag85a plasmid was confirmed in eukaryotic cells.Results: Fragments of 297 bp for tb10.4 and 1017 bp for ag85a were observed in agarose gel electrophoresis.Alignment of ag85a-tb10.4 genome sequence with reference genes in GenBank showed exact identities that indicate correction of all cloning procedures. Transfection of eukaryotic cells with pcDNA3.1+/tb10.4-ag85a vector and existence of tb10.4-ag85a fusion gene were both confirmed with RT-PCR.Conclusion: In this study, tb10.4 and ag85a genes were isolated from Mycobacterium tuberculosis H37Rv strain and cloned into pcDNA3.1+. Also, the capability of constructed vector in producing fusion ag85a-tb10.4 protein was confirmed with RT-PCR. pcDNA3.1+/tb10.4-ag85a vector can be used for further studies in future

The Iranian Plateau during the Bronze Age

The book compiles a portion of the contributions presented during the symposium “Urbanisation, commerce, subsistence and production during the third millennium BC on the Iranian Plateau”, which took place at the Maison de l’Orient et de la Méditerranée in Lyon, the 29-30 of April, 2014. The twenty papers assembled provide an overview of the recent archaeological research on this region of the Middle East during the Bronze Age. The socio-economic transformation from rural villages to towns and nations has prompted many questions into this evolution of urbanisation. What was the impact of interactions between cultures in the Iranian Plateau and the surrounding regions (Mesopotamia, the South Caucasus, Central Asia, Indus Valley)? What was the overall context during the Bronze Age on the Iranian Plateau? What was the extent and means of the expansion of the Kuro-Araxe culture? How did the Elamite Kingdom become established? What new knowledge has been contributed by the recent excavations and studies undertaken in the east of Iran? What was the influence of the Indus Valley culture, known as an epicentre of urbanisation in South Asia? What are the unique characteristics of the ancient cultures in Iran? While the urbanisation of early Mesopotamia has been the subject of much debate for several decades, this topic has only recently been raised in respect to the Iranian Plateau. This volume is the product of an international community from Iranian, European, and American institutions, consisting of recognised specialists in the archaeology of the Iranian Bronze Age. It provides an overview of the latest research, including abundant results from current on-going excavations. The current state of archaeological research in Iran, comprising many dynamic questions and perspectives, is presented here in the form of original contributions on the first emergence of towns in the Near and Middle East.L’ouvrage rassemble une partie des contributions présentées lors du colloque «Urbanisation, commerce, subsistance et production au iiie millénaire avant J.-C. sur le Plateau iranien» qui s’est tenu à la Maison de l’Orient et de la Méditerranée à Lyon les 29 et 30 avril 2014. Les vingt articles réunis livrent un état récent de la recherche archéologique dans cette région du Moyen‑Orient pour l’âge du Bronze. Le développement socio-économique entre le mode de vie rural et la formation des villes et des états soulève de nombreuses interrogations sur le processus de l’urbanisation. Quel est l’impact des relations culturelles entre le Plateau iranien et les régions adjacentes (Mésopotamie, Sud-Caucase, Asie centrale, vallée de l’Indus) ? Quel est le contexte global de l’âge du Bronze sur le Plateau Iranien ? Comment s’opère l’expansion de la culture Kuro-Araxe à partir du Caucase ? Comment le royaume élamite se met en place ? Quel est l’apport des fouilles et travaux récents dans l’Est iranien ? Quelle est l’influence de la vallée de l’Indus, un centre d’urbanisation important en Asie ? Comment se manifestent les singularités du monde iranien ? Alors que la thématique de l’urbanisation en Mésopotamie a été très débattue ces dernières décennies, cette question est abordée depuis peu pour le Plateau iranien. Le présent volume émane d’une communauté internationale d’archéologues d’institutions iraniennes, européennes et américaines, spécialistes reconnus de l’archéologie iranienne de l’âge du Bronze. Il dresse un panorama de l’état des recherches qui se nourrit amplement des travaux de terrain en cours. L’ouvrage rend compte de la dynamique actuelle de la recherche archéologique en Iran, riche de nouveaux questionnements et de nouvelles perspectives, et constitue un apport original à la réflexion sur l’émergence des villes au Moyen-Orient