ABSTRACT
Background: Causing site direct mutation can be one of the efficient methods to evaluate the characteristics and properties of various genes. Brucellosis is the most common zoonotic infectious disease that would cause great economic losses. Thus, recognition of pathogenic and immunogenic factors in the genus Brucella can lead to control this health problem
Objectives: Considering the importance of site direct mutation in identification of genome structure and numerous ways to achieve this goal, Overlap Extension PCR is introduced as an improved technique for the removal and replacement of the gene target
Methods: For this study, with two-step PCR using specific primers, upstream and downstream fragments from target gene and antibiotic resistance cassette from plasmid pET28a [+], were reproduced and were connected to each other. The resulting fragment was cloned in specific position of pBluescriptIISK[-] plasmid by the restriction enzymes. Then, the construction was transferred into the genome of Brucella abortus by electroporation method
Results: Fusion PCR product was obtained without any change in the nucleotide sequence and then it was cloned into pBluescriptIISK [-] plasmid, finally the construction was replaced and the target gene was deleted
Conclusions: The results of this study show that the Overlap Extension PCR is an optimized and modified technique to create mutations in the bacterial genome structure and can easily be used in the family Brucella
ABSTRACT
In present study the zoonotic role of cat in Bartonella henselae transmission have determined. It has done on 100 cats in 2 groups: indoor and outdoor and in 2 age's subgroups. Bartonella henselae was not isolated from blood culture of cats. 23 cats from 100 cats [23%] had antibodies against B. henselae. In this study there were no significant differences statistically in seroprevalence between cats and their owners [p<0.381]. Seroprevalence of cat owners was 18% and in control group [persons who own no cat] was 5%. There were significant differences [p<0.004] between cat owners and control group. Only 6 cats of 50 cats under 6 months old had antibodies to bartonella henselae, and in the other group 17 cats were seropositive and there were significant differences between these two groups [p<0.009] that showed seroprevalence in cats more than 6 months old is higher than the cats under 6 months old. 2 indoor cats from 50 indoor cats and 21 outdoor cats from 50 outdoor cats were seropositive and comparing of these two groups showed significant differences [p<0.0005], which confirmed indoor cats are less frequently infected than outdoor or stray cats