background: splicing by overlap extension (soe) pcr is used to create mutation in the coding sequence of an enzyme in order to study the role of specific residues in protein’s structure and function.objectives: we introduced a nested-soe-pcr (n –soe-pcr) in order to increase the specificity and generating mutations in a gene by soe-pcr.  materials and methods: genomic dna from bacillus thermoca...

Background: Splicing by overlap extension (SOE) PCR is used to create mutation in the coding sequence of an enzyme in order to study the role of specific residues in protein’s structure and function. Objectives: We introduced a nested-SOE-PCR (N –SOE-PCR) in order to increase the specificity and generating mutations in a gene by SOE-PCR.   Materials and Methods: Genomic DNA from Bacillus thermo...

Background: Rotavirus group A (RVA) is recognized as a major cause of severe gastroenteritis in children and new-born animals. Nonstructural protein 4 (NSP4) is responsible for the enterotoxic activity of these viruses in the villus epithelial cells. Amino acids 114-135 of NSP4 are known to form the diarrhea-inducing region of this viral enterotoxin. Therefore, developing an NSP4 lacking the en...

Epitope tagging is a method of expressing proteins whereby an epitope for a specific monoclonal antibody is fused to a target protein using recombinant DNA techniques. The aim of this study was to sub-clone the peroxisomal protein (PEP) cDNA into a mammalian expression vector leading to the formation of a  chimeric PEP-cDNA containing the FLAG epitope. The FLAG-PEP recombinant cDNA was construc...

Background: In reverse genetics studies, minigenomes of negative sense RNA viruses are used to model the process virus replication and evaluate rescue system in course construction recombinant virus. current study, a minigenome (MG) for lentogenic D58 vaccine strain Avian orthoavulavirus 1 (AOAV-1) (Newcastle disease virus) was developed establish genetic this strain. Methods: The constructed b...

Four exons of the human genomic GM-CSF gene were assembled together using gene splicing by overlap extension (SOE) method. The resulting nucleotide sequence was cloned in the pET23a(+) ex‌pression vector under the control of strong bacteriophage T7 transcription and translation signals. The construct obtained was Transferred into the E. coli strain, BL21(DE3) pLysS and IPTG was used for inducti...

four exons of the human genomic gm-csf gene were assembled together using gene splicing by overlap extension (soe) method. the resulting nucleotide sequence was cloned in the pet23a(+) ex pression vector under the control of strong bacteriophage t7 transcription and translation signals. the construct obtained was transferred into the e. coli strain, bl21(de3) plyss and iptg was used for inducti...