shohre zare

Enterotoxigenic Escherichia coli (ETEC) and Enterohemorrhagic Escherichia coli O157:H7 , the most common strains,  are causes diarrhea that can kill  hundreds of thousands of children annually. The development of an effective combination vaccine for enterohemorrhagic Escherichia coli and Enterotoxigenic Escherichia coli is very important.

In this study, the sicl gene was amplified and Subcloned as a DNA vaccine. The sequence of antigen encoding genes was evaluated from the genebank and their epitopes were evaluated to design of primer for the synthetic chimeric gene and was amplified by PCR. Subcloning of  a multipartal chimeric gene in eukaryotic expression vector was performed to make a DNA vaccine and finally the protein was purified by nickel chromatography and evaluated by Western blotting.

The immunoblotting results of the expression of SICL chimeric protein indicated the presence of a 76 kDa band in the form of insoluble particles after 12 hours induction. Purification of the recombinant protein using His tag sequencee and confirmation of the purified protein with the recombinant protein specific antibody demonstrated the accuracy of the protein expression.

Protein expression, purification and verify by western blotting showed that this recombinant chimeric protein (SICL) can be expressed in eukaryotic host.

Archaeogenetics, a nascent and emerging science, is based on a combination of biology and genetics which has a high potential in discovering the untold facts about livelihood and culture of ancient times, such as modern human origins and diets evolutionary trend. This area of biology, also focus on migration and geographical distribution of humans and genetic variation in ancient DNA samples around the world in comparison to modern humans. Highly polymorphic DNA regions like STR found on the male-specific Y chromosome in nucleus and mitochondrial (mt) DNA are used in archaeogenetics to study genetic variation within a target population. mtDNA D-loop region is very polymorphic that consists of two hyper variable regions including HVI and HVII with a large variety in different human populations. Analysis on these regions of mtDNA using ancient excavated human bones will lead to determine the genetic composition of human mtDNA known as haplogroups. mtDNA can be used to identify the ancient ethnic groups, trace descendants of ancestors and their migration trails.This is achieved by comparing mtDNA haplogroups between different ethnic groups all over the world.In this study, we have performed analysis using ancient DNA extracted from 5 excavated human bones. Ancient human bones were obtained from Haft-Tepe. We performed PCR amplifications for HVI and HVII regions of the mtDNA followed by sequencing with ABI. Then comparisons The Cambridge Reference Sequence and the sequences obtained in the current study were done at NCBI site. Hoplogroup residents were R2 and R5. By tracking haplogroups from indo-Iranian , we can conclude that these haplogroups dated to 5000 B.P in Iran, and possibly originated in southwestern Iran and from the Elamite civilization.

One of the interdisciplinary approaches which recently had a key role in resolving of archaeological issues, especially domestication process, was Archaeogenetics. Caprines were domesticated in the early Neolithic period and their domestication was a major part of the process that led to the invention of agriculture. Since goat has had a key role in subsistence economy of human societies in different environments hence it became one of the main aims of archeogentics studies. By use of aDNA of goats and archaeogenetics approaches, this paper extracted and analyzed 10 samples of goat aDNA from Neolithic period of Kashan and Qazvin plains and determined their phylogenetic relations with modern domestic and wild goats to shed more light on the life of late Neolithic society of central plateau of Iran. Results show that these samples have closest genetic relations with haplogroup A of wild goats which now live in southeastern of Turkey. Hence archeogenetic evidences show that by 7500 B.P, Neolithic societies of Iran central plateau used goats which were domesticated about 10000 B.P in southeastern Turkey.