IN VIVO EXPRESSION OF IDENTIFIED HEPATITIS C VIRUS GENI (S) FOR DIAGNOSTIC PURPOSES
Loaa Abdala Tag Eldeen;
Abstract
Hepatitis c virus (HCV) ha...; hl'l'll iJcntilicd as the m jilr C;\IJ .JiiH• a,gent or rarcntl'r
encoding a full length of the HCV genotype 4 core protein \Vas amplilled from HCV
infected human serum by H.T-PCR using the synthetic oligonucleotied primers designed for amplification of the open reading frame coding for the core region. The RNA was used to generate eDNA using Core int. antisense primer in a separate step, then eDNA was used for nested PCR using Core ext. sense and Core int. antisense followed by amplification with primers, Core int. sense and Core int. antisense The amplified core segment was cloned into pTARGET mammalian expression vector to generate pTARGET-HCYCore plasmid. The construct was transformed in its host £.Coli bacterial strain and analyzed for the presence and ori• ntatinn of the in...,c n. The pTARC:FT-HCVCorc plasmid was subjected to super-scaling and purified by polycthylinglycol (PEG). lo group of llllCC are injected intramuscular with the plasmid-core construct. The first group wns injected twice by 120).lg of the construct with 5 day interval and sacrificed in the 6111 day and their muscle homo cnatc supanatanl wa; suhjcctcJ to wcsh:ril blouing. The second group was primed by 120g of construct and boosted twice with 60 g of the construct at 0, 3 and 5 week intervals. After 7 weeks thoir blood were collected and tested for the presence of induced anti-core antibody. Using recombinant DNA construct represent a new technique for production of polyclonal antibodies against HCV-corc antigen.
encoding a full length of the HCV genotype 4 core protein \Vas amplilled from HCV
infected human serum by H.T-PCR using the synthetic oligonucleotied primers designed for amplification of the open reading frame coding for the core region. The RNA was used to generate eDNA using Core int. antisense primer in a separate step, then eDNA was used for nested PCR using Core ext. sense and Core int. antisense followed by amplification with primers, Core int. sense and Core int. antisense The amplified core segment was cloned into pTARGET mammalian expression vector to generate pTARGET-HCYCore plasmid. The construct was transformed in its host £.Coli bacterial strain and analyzed for the presence and ori• ntatinn of the in...,c n. The pTARC:FT-HCVCorc plasmid was subjected to super-scaling and purified by polycthylinglycol (PEG). lo group of llllCC are injected intramuscular with the plasmid-core construct. The first group wns injected twice by 120).lg of the construct with 5 day interval and sacrificed in the 6111 day and their muscle homo cnatc supanatanl wa; suhjcctcJ to wcsh:ril blouing. The second group was primed by 120g of construct and boosted twice with 60 g of the construct at 0, 3 and 5 week intervals. After 7 weeks thoir blood were collected and tested for the presence of induced anti-core antibody. Using recombinant DNA construct represent a new technique for production of polyclonal antibodies against HCV-corc antigen.
Other data
| Title | IN VIVO EXPRESSION OF IDENTIFIED HEPATITIS C VIRUS GENI (S) FOR DIAGNOSTIC PURPOSES | Other Titles | التعبير في الح لجين ( جينات ) معين لفيروس الالتهاب الكبدي ج للأغراض التشخيصية | Authors | Loaa Abdala Tag Eldeen | Issue Date | 2004 |
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