RESUMO
Background: Hepatitis viruses are one of the serious medical problems and Hepatitis B is one of the chief transferable disease via blood and its products. Nowadays, 11 genotypes of hepatitis B have known over the world by the genome sequencing. Hepatitis B viruses have special geographical distribution. The clinical importance of hepatitis B viruses and its relation with the mutations has recognized. The purpose of this study was to check the presence and prevalence of Hepatitis B virus genotypes among the referrals attended to the medical diagnostic laboratories in Kerman province
Materials and Methods: In this cross-sectional study, twenty-one specimens were collected from blood samples available in the medical diagnostic laboratories of Kerman province during one year. After DNA extraction, PCR was carried on by specific primers, then they were sequenced. The obtained sequenced were compared with sequences in the NCBI gene bank and blasted for identification of their genotypes
Results: Seven samples from twenty one samples [33.3 percent] had D genotype, 13 samples from 21 [62 percent] had D3 subgenotype and 1 sample from 21 [4.7 percent] had D4 subgenotype
Conclusion: The prevalence of these genotypes in the Kermanian patients that recognized in this study can help to provide diagnostic kits for hepatitis B virus
RESUMO
Background: The homologous recombination [HR] is one of the conventional cloning methods for the production of recombinant DNA. It is a quick method for in vivo DNA cloning without using the high cost restriction enzymes. A few modifications in the cloning procedure can increase the efficiency of this method.
Materials and Methods: In this study, effect of heating on the rate of the IgG1 heavy chain gene cloning was investigated in the HR method and then it was compared with HR method without heating and traditional cloning method. For doing this comparison, three cloning methods including HR, HR with the heat treatment, and traditional cloning were used to clone the human IgG1 heavy chain into the pcDNA3.1[+] plasmid.
Results: The results showed that adding heat in the HR method converts insert and vector from the double strand DNA to the single strand DNA. This allows them to copulate with each other better and faster than the two other methods. The heat addition also helps the cell enzyme system for a faster and easier recombination and moreover it increases the cloning efficiency of the HR method in case of in vitro processing.
Conclusion: The results showed that giving heat in the HR method increases cloning rate 7.5% and this increase reaches 10% in comparison with the traditional method.