جستجوی مقالات مرتبط با کلیدواژه "coat protein" در نشریات گروه "بیوتکنولوژی و ژنتیک گیاهی"

The use of plant viruses as carriers of anti-cancer drugs in medicine is increasing day after day. One of the most important potato viruses is Potato X Potexvirus (PVX) which is globally expanded and is one of the first plant viruses used as a carrier of anticancer drugs. In this research, in order to produce PVX particles, its Coat protein (CP) gene was expressed. First, the RT-PCR reaction was performed for CP amplification using specific primers containing the endonuclease digestion sites. Electrophoresis of the PCR product on agarose gel confirmed the amplified fragment with a length of 714 bp. Then the CP gene and pGBKT7 plasmid were digested by BamHI and EcoRI enzymes and after purification, ligation was done between them. Recombinant plasmid pGBKT7-CPPVX was transferred to E. coli bacteria by electroporation. After selecting the bacteria containing the recombinant plasmid by PCR and extracting the plasmids from them, sequencing confirmed the correctness of the cloned gene sequence. Then, the CP gene was cloned into pET-21a expression plasmid by digestion and ligation method. The recombinant plasmid was transferred to BL21 strain of E. coli and cultured. By adding IPTG in the bacterial culture medium, CP protein was expressed from the plasmid and bacterial proteins were extracted. Protein electrophoresis showed the expression of CP. After purification, this protein can be used as a carrier of anti-cancer drugs.

Viruses are the amazing particles in nature that is designed to reproduce with a minimal genetic set. This vital process is very complex and requires the time and coordinated activities of a number of virus and host proteins. In order to the genome packaging, the virus protein subunits interact with each other and with viral nucleic acids. The general principle in packaging is the interaction between the structure of coat protein and a specific signal in RNA. Replication of +ssRNA viruses and their nucleocapsid assembly occurs in the cytoplasm and requires Pro-Pro and Pro-RNA interactions. The packaging process in them is spontaneous and does not require ATP. The interaction between the coat protein subunits and the genomic RNA is controlled by a unique structure in the sequence that is highly desired to interaction with the coat protein. The presence of these unique structures in the viral nucleic acid, called the packaging signal or OAS (origin of assembly sequence), causes viral nucleic acid to be distinguished from other cellular RNA molecules in the region which assembly takes place. General knowledge of the precise mechanisms of viruses, the configuration and aggregation of viruses, and the packing of their genomes that lead to the formation of stable viral structures is an important prerequisite for understanding the overall biology of viruses.